Epinephrine formulations

ABSTRACT

Pharmaceutical compositions comprising epinephrine, methods of administration, and methods of making the same. Compositions may comprise at least one of an active agent, a pH raising agent, an antioxidant, a transition metal complexing agent, a pH lowering agent, a tonicity regulating agent, optionally a preservative, and optionally a solvent.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/817,253, filed Mar. 12, 2020, which is incorporated by referenceherein in its entirety, which is a Continuation of U.S. patentapplication Ser. No. 16/258,204, filed Jan. 25, 2019, which is aContinuation of U.S. patent application Ser. No. 15/969,500, filed May2, 2018, which is a Continuation of U.S. patent application Ser. No.15/455,991, filed Mar. 10, 2017, which is a Continuation-in-Part of U.S.patent application Ser. No. 14/863,112, filed Sep. 23, 2015, now U.S.Pat. No. 10,130,592, issued Nov. 20, 2018, which is a Continuation ofU.S. patent application Ser. No. 14/818,121, filed Aug. 4, 2015, nowU.S. Pat. No. 9,295,657, issued Mar. 29, 2016, which is a Continuationof U.S. patent application Ser. No. 14/657,990, filed Mar. 13, 2015, nowU.S. Pat. No. 9,119,876, issued Sep. 1, 2015. The disclosures of theprior applications are hereby incorporated in their entirety byreference.

FIELD OF THE INVENTION

The present invention provides pharmaceutical compositions comprisingepinephrine, methods of administration, and methods of making the same.

BACKGROUND

Epinephrine, also known as4-[(1R)-1-Hydroxy-2-(methylamino)ethyl]-1,2-benzenediol, is the activeprinciple of the adrenal medulla and an endogenous catecholamine whichacts directly on both alpha and beta adrenergic receptors. When used inpharmaceutical compositions, epinephrine can act as a non-selectivealpha and beta adrenergic agonist and can work rapidly to improvebreathing, stimulate the heart, raise dropping blood pressure, reversehives, and reduce swelling of the face, lips, and throat. Uses forepinephrine include emergency treatment of allergic reactions (Type 1),including anaphylaxis, induction and maintenance of mydriasis duringintraocular surgery, treatment of bronchospasm, sensitivity reactions,cardiac arrhythmias, GI and renal hemorrhage, superficial bleeding,premature labor, hypoglycemia, and cardiogenic, hemorrhagic, andtraumatic shock. Epinephrine can also be used to increase blood flow inACLS during CPR, as an adjunct to local anesthesia, and for radiographicuses.

One example of a pharmaceutical composition with epinephrine as anactive agent is Adrenalin®, a sympathomimetic agent manufactured anddistributed by Par Sterile Products, LLC. Adrenalin® is an epinephrineinjection suitable for subcutaneous, intracameral, and intramuscularadministration. When diluted, it may also be administered as ophthalmicirrigation or intracameral injection. Adrenalin® is used primarily as anemergency treatment of allergic reactions (Type 1), includinganaphylaxis, and for induction and maintenance of mydriasis duringintraocular surgery.

Adrenalin® is a clear, colorless solution containing 1 mg/mL epinephrinein a clear glass vial. Adrenalin® is currently available in 1 mLsingle-dose and 30 mL multi-dose formulations. For the 1 mL product,each 1 mL of Adrenalin® contains 1 mg epinephrine, 9.0 mg sodiumchloride, 1.0 mg sodium metabisulfite, hydrochloric acid to adjust pH,and water for injection. For the 30 mL product, each 1 mL of Adrenalinesolution contains 1 mg epinephrine, 9.0 mg sodium chloride, 1.5 mgsodium metabisulfite, hydrochloric acid to adjust pH, 5.4 mgchlorobutanol as a preservative and water for injection. The pH range ofAdrenalin® is 2.2-5.0.

Adrenaline 1 mL is FDA approved for an 18 month shelf life. TheAdrenalin® 30 mL product is approved for an even shorter shelf life of14 months. Shelf life is limited by the formation of degradants, whichmainly comprise epinephrine sulfonic acid (ESA) and D-epinephrine, anenantiomer of L-epinephrine that has insignificant therapeutic activity.The currently approved Adrenalin® 1 mL product has a total impuritylimit of ≤20%. Adrenalin® 1 mL has a concentration limit of ≤13.5% ESAand ≤9.5% D-epinephrine. The currently approved Adrenaline 30 mL producthas a total impurity limit of ≤20%. Adrenalin®30 mL has a concentrationlimit of ≤14.5% ESA and ≤9.5% D-epinephrine.

There is currently a need in the art for improved epinephrine-containingpharmaceuticals. It is an object of the present invention to provide anepinephrine-containing pharmaceutical composition that addresses some ofthe limitations of present formulations.

SUMMARY OF THE INVENTION

The present invention provides a pharmaceutical composition comprisingepinephrine. The composition may be useful for emergency treatment ofallergic reactions (Type 1), including anaphylaxis, induction andmaintenance of mydriasis during intraocular surgery, treatment ofbronchospasm, sensitivity reactions, cardiac arrhythmias, GI and renalhemorrhage, superficial bleeding, premature labor, hypoglycemia, andcardiogenic, hemorrhagic, and traumatic shock. The composition may alsobe used to increase blood flow in ACLS during CPR, as an adjunct tolocal anesthesia, and for radiographic uses. The composition may beadministrable via subcutaneous, intracameral, intravenous, andintramuscular injection, infusion, intra-arterial administration,intracardiac injection, endotracheal administration, intraosseousadministration, oral inhalation, topical administration, and asophthalmic irrigation.

The present invention also provides for a composition comprising atleast one of an active agent, a pH raising agent, an antioxidant, atransition metal complexing agent, a pH lowering agent, a tonicityregulating agent, optionally a preservative, and optionally a solvent.

The present invention also provides for compositions with low levels ofD-epinephrine.

The present invention also provides for compositions with low levels ofepinephrine sulfonic acid (ESA).

The present invention also provides for compositions with low levels ofoxidative degradants.

The present invention also provides for compositions with low levels oftotal impurities.

The present invention also provides for compositions comprising a pHraising agent.

The present invention also provides for compositions comprising anantioxidant.

The present invention also provides for compositions that may resistsignificant pH change over shelf life.

The present invention also provides for a method of making suchcompositions.

The present invention also provides for a method of treating a medicalcondition comprising administering to a subject in need thereof acomposition of the present invention.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a chromatogram of the Impurities Marker Solution usingisocratic HILIC with 15% water.

FIG. 2 is a chromatogram of the Impurities Marker Solution usingisocratic HILIC with 18% water.

FIG. 3 is a chromatogram of the Impurities Marker Solution usingisocratic HILIC with 22% water.

FIG. 4 is a chromatogram of the Impurities Marker Solution usinggradient HILIC separation.

FIG. 5 shows the total concentration of impurities at various timepoints during the study according to Example 25.

FIG. 6 shows the total concentration of impurities at various timepoints during the study according to Example 26.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a pharmaceutical composition comprisingepinephrine. The composition may be useful for emergency treatment ofallergic reactions (Type 1), including anaphylaxis, induction andmaintenance of mydriasis during intraocular surgery, treatment ofbronchospasm, sensitivity reactions, cardiac arrhythmias, GI and renalhemorrhage, superficial bleeding, premature labor, hypoglycemia, andcardiogenic, hemorrhagic, and traumatic shock. The composition may alsobe used to increase blood flow in ACLS during CPR, as an adjunct tolocal anesthesia, and for radiographic uses. The composition may beadministrable via subcutaneous, intracameral, intravenous, andintramuscular injection, infusion, intra-arterial administration,intracardiac injection, endotracheal administration, intraosseousadministration, oral inhalation, topical administration, and asophthalmic irrigation.

The present invention provides for a composition comprising at least oneof an active agent, a pH raising agent, an antioxidant, a transitionmetal complexing agent, a pH lowering agent, a tonicity regulatingagent, optionally a preservative, and optionally a solvent.

In some preferred embodiments, the composition may comprise an activeagent, a pH raising agent, an antioxidant, a pH lowering agent, and atonicity regulating agent. In some preferred embodiments, thecomposition may comprise an active agent, an antioxidant, and a tonicityregulating agent. In some preferred embodiments, the composition maycomprise an active agent, a transition metal complexing agent, a pHraising agent, a pH lowering agent, and a tonicity regulating agent. Insome preferred embodiments, the composition may comprise an activeagent, a transition metal complexing agent, an antioxidant, and atonicity regulating agent. In some preferred embodiments, thecomposition may comprise an active agent, a pH raising agent, a pHlowering agent, and a tonicity regulating agent. One of ordinary skillwill appreciate that the combination of components in the compositionmay be varied in order to obtain compositions with desired properties.

In some embodiments, the active agent comprises epinephrine and/or saltsthereof. Examples of epinephrine salts include, but are not limited to,acetate, carbonate, citrate, hydrochloride, hydrocyanide, hydrofluoride,nitrate, nitrite, phosphate, and sulfate salts. Preferably, the salt isa hydrochloride salt.

In some embodiments, the active agent is present at a concentrationsufficient for any of the uses described herein. In some embodiments,the active agent is present at a concentration of about 0.01 to 2 mg/mL,preferably about 0.01 to 1 mg/mL, more preferably about 0.01 to 0.5mg/mL, more preferably about 0.01 to 0.3 mg/mL, and most preferably 0.01to 0.2 mg/mL. Alternatively, the active agent is present at aconcentration of about 0.1 to 2 mg/mL, preferably about 0.5 to 1.5mg/mL, more preferably about 0.75 to 1.25 mg/mL, and most preferablyabout 1 mg/mL. In some embodiments, the active agent is present at aconcentration of 0.114 mg/mL or 1.14 mg/mL.

The present invention provides for a composition that may comprise atleast one pH raising agent in an amount that raises the pH of thecomposition. In some embodiments, the pH raising agent comprises abuffer system. The term “buffer system” refers to a component present ina composition or solution which may provide a resistance to significantchange in pH caused by a strong acid or base. A buffer system maycomprise a single agent or more than one agent, such as a weak acid andits conjugate base. A buffer system may provide a resistance to asignificant pH change by interacting with a strong acid or strong basein a composition or solution, thereby at least partially preventing thepH of the composition or solution from changing significantly.

Generally, a buffer system has one or more buffer ranges wherein thebuffer system has the ability to provide resistance to significant pHchange. When a composition or solution comprising the buffer system hasa pH inside the buffer system's buffer range, the pH of the compositionor solution will not change significantly with the addition of equimolaramounts of a strong acid or strong base.

The buffer range of a buffer system is related to the acid dissociationconstant (K_(a)) of one or more weak acids comprised by the buffersystem. The term “acid dissociation constant” refers to the equilibriumconstant of a dissociation reaction of an acid. The midpoint of a bufferrange for a buffer system is generally about the logarithmic measure ofthe acid dissociation constant (i.e., the pK_(a), equal to −log₁₀K_(a))of a weak acid comprised by the buffer system.

In some embodiments, the pH raising agent may comprise one or morepK_(a) values. In some embodiments, the pH raising agent may comprise apK_(a) value that is within +/−1 unit of the initial pH of thepharmaceutical composition in the form of a solution without the pHraising agent. In some embodiments, the pH raising agent may have apK_(a) value within the range of about 2 to 5, preferably within therange of about 3 to 4.5, more preferably within the range of about 3.5to 4.5, and most preferably about 4.

In some embodiments, the pH raising agent may have a buffer range from apH of about 2 to 5, preferably from a pH of about 3 to 4.5, and mostpreferably from a pH of about 3.5 to 4.5.

In some embodiments, the pH raising agent may provide a resistance to asignificant change in pH that other components of the composition mayotherwise cause in the absence of the pH raising agent. In somepreferred embodiments, the pH raising agent may provide a resistance tothe composition's pH being significantly lowered by degradants and/orreaction products of other components of the composition. For example,the pH raising agent may help resist an increase in acidity associatedwith the chemical reaction of an antioxidant.

In some embodiments, the pH raising agent may be present at aconcentration in the range of about 0.1 and 4 mg/mL, preferably in therange of about 1 and 3 mg/mL, preferably in the range of about 1.5 and2.5 mg/mL, and most preferably about 2.25 mg/mL. In some embodiments,the pH raising agent may be present at a concentration that provides aresistance to a significant change in the composition's pH despitephysical and chemical changes of the composition after a certain periodof shelf life.

In some embodiments, the pH raising agent is selected such that thedegradation of other components of the composition is slowed or reduced,as compared to a composition in which the pH raising agent is notpresent. For example, the pH raising agent may at least in part helpprevent significant degradation of an antioxidant after a certain periodof shelf life.

In some embodiments, the pH raising agent is selected such that lessantioxidant is present, as compared to a composition in which the pHraising agent is not present.

For example, the pH raising agent may be selected such that theconcentration of antioxidant necessary for preventing and/or inhibitingthe formation of unacceptable amounts of oxidative degradants in thecomposition after a certain period of shelf life is lowered by in therange of about 10% to 90%, preferably in the range of about 20% to 80%,more preferably in the range of about 25% and 75%, more preferably inthe range of about 30% and 70%, and most preferably in the range ofabout 40% and 60%, as compared to a composition in which the pH raisingagent is not present.

In some embodiments, the pH raising agent and pH raising agentconcentration is such that compositions conform to the limit on totalacidity imposed by the United States Pharmacopeia (USP) 37-NF 32, S2,monograph, effective from Dec. 1, 2014. For example, the pH raisingagent may be selected such that when 5.0 mL of the composition istitrated with 0.01 N NaOH, the total amount of titrant necessary toreach a pH of 7.4 is no more than 25 mL. The pH raising agent may beselected such that the composition has a titratable acid concentrationof in the range of about 30 to 70 mM, preferably in the range of about40 and 60 mM, and most preferably about 50 mM.

In some embodiments, the pH raising agent is suitable for subcutaneous,intracameral, intramuscular, parenteral, and/or ophthalmic use.

Preferably, pH raising agents comprise the acids or salt forms of one ormore of lactate, tartarate, glutamate, malate, citrate, gluconate,benzoate, succinate, acetate, glycine, and aspartate, as well as lithiumhydroxide, sodium hydroxide, potassium hydroxide, rubidium hydroxide,cesium hydroxide, calcium hydroxide, strontium hydroxide, and bariumhydroxide. Preferably, the pH raising agent comprises at least one oftartaric acid and sodium hydroxide, and more preferably both of thesecompounds.

The present invention provides for a composition that may comprise atleast one antioxidant. The term “antioxidant” refers to a component in acomposition that may prevent and/or inhibit the formation ofunacceptable amounts of oxidative degradants in the composition after acertain period of shelf life. In some embodiments, the antioxidant mayreact with oxygen that might otherwise compromise the composition byproducing impurities in the composition. Oxygen may originate from thecomposition's environment or the composition itself. For example, oxygenmay originate from residual oxygen present in the headspace of vialscontaining the composition.

In some embodiments, the oxidative degradants comprise one or more ofadrenalone, adrenochrome, adrenolutin, adrenochrome sulfonate, andmelanins.

In some embodiments, the antioxidant may limit the formation ofoxidative degradants in the composition to less than about 1%,preferably less than about 0.5%, more preferably less than about 0.1%,more preferably less than about 0.05%, more preferably less than about0.04%, more preferably less than about 0.03%, more preferably less thanabout 0.02%, more preferably less than about 0.01%, most preferably lessthan about 0.009%, after a certain period of shelf life.

In some embodiments, the antioxidant may inhibit and/or prevent thecomposition from undergoing unacceptable physical changes. In someembodiments, the antioxidant may inhibit and/or prevent unacceptablecomposition color change. In some embodiments, the antioxidant mayinhibit and/or prevent unacceptable amounts of insoluble particles.

In some embodiments, the antioxidant may inhibit and/or prevent thecomposition from undergoing unacceptable color changes that may bedetected by visually examining a portion of a sample in a clear, glasstest tube against a white background. In some embodiments, theantioxidant may inhibit and/or prevent the composition from undergoingunacceptable color changes that may be detected by comparing theabsorbance of a sample of the composition in a 1-cm quartz cell using aspectrophotometer set at 460 nm with a 0.0004 N Iodine Standard Solutionthat is prepared with water and iodine. In some embodiments, theantioxidant may inhibit and/or prevent the composition from containingunacceptable amounts of particulate matter that may be detected by themethods described in USP <788> and/or USP <789>, the disclosures ofwhich are included herein by reference.

In some embodiments, the antioxidant may be present at the lowestconcentration that will inhibit and/or prevent the composition fromundergoing unacceptable physical changes. In some embodiments, theantioxidant may be present at the lowest concentration that will inhibitand/or prevent unacceptable oxidation of components of the composition.

In some embodiments, the antioxidant may be present at a concentrationof in the range of about 0.1 to 3.0 mg/mL, preferably in the range ofabout 0.3 to 2.7 mg/mL, more preferably in the range of about 0.5 to 2.5mg/mL, more preferably in the range of about 0.7 to 2.3 mg/mL, morepreferably in the range of about 1.0 and 2.0 mg/mL, more preferably inthe range of about 1.3 and 1.7 mg/mL, most preferably about 1.5 mg/mL.In some embodiments, the antioxidant may be present at a concentrationof in the range of about 0.1 to 1.1 mg/mL, preferably in the range ofabout 0.1 to 0.9 mg/mL, more preferably in the range of about 0.2 and0.8 mg/mL, more preferably in the range of about 0.3 and 0.7 mg/mL, morepreferably in the range of about 0.4 and 0.6 mg/mL, and most preferablyabout 0.4 to 0.5 mg/mL.

In some embodiments, the concentration of antioxidant in the compositionafter a certain period of shelf life may be affected by other componentsof the composition that reduce and/or slow the rate of antioxidantdegradation.

In some embodiments, other components of the composition may reduceand/or slow oxidation of the antioxidant after a certain period of shelflife. For example, when the antioxidant is sodium bisulfite, sodiummetabisulfite, or another sulfite, other components of the compositionmay reduce and/or slow the oxidation of bisulfite to bisulfate.

In some embodiments, other components of the composition may reduceand/or slow protonation and/or desolvation reactions involving theantioxidant over a certain period of shelf life. For example, when theantioxidant is sodium bisulfite, sodium metabisulfite, or anothersulfite, other components of the composition may reduce and/or slow theconversion to sulfur dioxide.

In some embodiments, other components of the composition may reduceand/or slow the degradation of antioxidant by raising the composition'sinitial pH and/or the pH of the composition after a certain period ofshelf life, as compared to a composition in which the other componentsare not present. In some embodiments, the pH raising agent may, at leastin part, lead to a reduction in the total degradation and/or degradationrate of antioxidant in the composition after a certain period of shelflife.

In some embodiments, other components are present in the composition ata concentration that reduces the concentration of antioxidantdegradation products present in the composition after a certain periodof shelf life. In some embodiments, other components are present in thecomposition at a concentration that increases the concentration of theantioxidant present in the composition after a certain period of shelflife, as compared to a composition in which the other components are notpresent. In some embodiments, the reduction and/or inhibition ofunacceptable antioxidant degradation after a certain period of shelflife may reduce the initial concentration of antioxidant necessary toinhibit and/or prevent the composition from undergoing unacceptablephysical changes after a certain period of shelf life. In someembodiments, less than about 90% of the initial amount of antioxidant isdegraded after a certain period of shelf life, more preferably less thanabout 85%, more preferably less than about 80%, more preferably lessthan about 75%, more preferably less than about 70%, more preferablyless than about 60%, more preferably less than about 50%, morepreferably less than about 40%, more preferably less than about 30%,more preferably less than about 20%, and most preferably less than about10%.

Preferably, pharmaceutically-acceptable antioxidants comprise one ormore of an amino acid sulfite (e.g L-lysine sulfite), ascorbic acid,ascorbyl palmitate, benzotriazol, butylhydroxyanisole (BHA),butylhydroxytoluene (BHT), citric acid, cysteine, cysteinehydrochloride, disodium calcium ethylenediaminetetraacetate, disodiumethylenediaminetetraacetate, dithiothreitol, DL-alpha-tocopherol,erythorbic acid, ethoxyquin, ethylenediaminetetraacetic acid salts,fumaric acid, glutathione, guaiac, homocysteine, isopropyl citrate,N-acetylcysteine, mercaptoethane sulfonic acid, L-ascorbate stearateesters, monothioglycerol, nordihydroguaiaretic acid (NDGA), palmiticacid ascorbic acid,pentaerythrityl-tetrakis[3-(3,5-di-t-butyl-4-hydroxyphenyl)propionate]2-mercaptobenzimidazole,cysteine ethyl ester, dithiothreitol, potassium dichloroisocyanurate,propyl gallate, rongalite (CH2OHSO2Na), sodium ascorbate, sodiumbisulfite, sodium edetate, sodium erythorbate, sodium hydrogen sulfite,sodium metabisulfite, sodium pyrosulfite 1,3-butylene glycol, sodiumsulfite, methionine, thioglycolic acid, sodium thioglycolate, sodiumthiosulfate, soybean lecithin, tert-butyl hydroquinone, thioglycerol,thiourea, TPGS (tocopherol polyethylene glycol succinate), vitamin E andderivatives thereof, α-thioglycerin, and/or salts thereof. Preferably,the antioxidant comprises sodium bisulfite or sodium metabisulfite. Inother preferred embodiments, the composition is free of sulfites such assodium bisulfite or sodium metabisulfite.

According to some preferred embodiments, the antioxidant may comprise asulfur-containing organic compound. Preferably, the sulfur-containingorganic compound is a thiol-containing organic compound. As used herein,the term “organic compound” refers to a compound belonging to the classof compounds characterized by their inclusion of carbon atoms in theirmolecular backbone. The term “thiol” refers to a carbon-bondedsulfhydryl group. Examples of sulfur-containing organic compoundssuitable for use in accordance with the invention include, but are notlimited to, cysteine, N-acetylcysteine, glutathione, methionine,monothioglycerol, cysteine ethyl ester, thioglycolic acid, thiolacticacid, dithiothreitol, reduced glutathione, mercaptoethane sulfonic acid,and salts thereof.

According to some aspects, the composition may have an initial activeagent overage that is reduced when compared with a composition whereinthe antioxidant is not present and/or is different than the antioxidantof the present invention. As used herein, “overage” refers to the amountof a component present in a composition that is more than (or “over”)the amount indicated on the label for the composition. For example,traditional epinephrine-containing compositions may have an overage ofup to 25% of the amount indicated on the label. Indeed, the USP 39monograph for epinephrine injections in water requires a formulation tocontain no less than 90.0% and no more than 115.0% of the labeled amountof epinephrine. That is, the USP 39 monograph allows for up to a 25%difference between a label claim and the actual amount.

According to some aspects, the present composition may have an initialactive agent overage of less than about 20%, preferably less than about19%, more preferably less than about 18%, more preferably less thanabout 17%, more preferably less than about 16%, more preferably lessthan about 15%, more preferably less than about 14%, more preferablyless than about 13%, more preferably less than about 12%, morepreferably less than about 11%, more preferably less than about 10%,more preferably less than about 9%, more preferably less than about 8%,more preferably less than about 7%, more preferably less than about 6%,more preferably less than about 5%, more preferably less than about 4%,more preferably less than about 3%, more preferably less than about 2%,more preferably less than about 1%, and most preferably about 0%.

The present invention provides a composition that may include one ormore transition metal complexing agents.

In some embodiments, the transition metal complexing agent may be achelating agent. The term “chelating agent” refers to a substancecapable of chelation, i.e, the formation or presence of two or moreseparate coordinate bonds between a polydentate ligand and a singlecentral atom.

In some embodiments, the transition metal complexing agent may reducethe catalytic activity of trace metals present in the composition. Insome embodiments, the transition metal complexing agent may chelatetrace metals in the composition that may otherwise increase and/oraccelerate the degradation of components in the composition. Examples oftrace metals include, but are not limited to, iron, magnesium, lithium,zinc, copper, chromium, nickel, cobalt, vanadium, arsenic, molybdenum,manganese, selenium, and calcium.

In some embodiments, the transition metal complexing agent may chelatetrace metals in the composition that may otherwise increase and/oraccelerate degradant formation in the composition. In some embodiments,the transition metal complexing agent may inhibit the formation ofdegradants formed from the interaction of epinephrine, bisulfite, andoxygen. In some embodiments, these degradants comprise one or more ofImpurity A, Impurity B, and Unknown C.

In some embodiments, the concentration of the transition metalcomplexing agent present in the composition may be such that theconcentration of Impurity A, Impurity B, or Unknown C is about 1% orless of the composition after a certain period of shelf life, preferablyabout 0.9% or less, more preferably about 0.8% or less, more preferablyabout 0.7% or less, more preferably about 0.6% or less, more preferablyabout 0.5% or less, more preferably about 0.4% or less, more preferablyabout 0.3% or less, more preferably about 0.2% or less, and mostpreferably about 0.1% or less.

In some embodiments, the transition metal complexing agent may bepresent at the lowest concentration that reduces and/or inhibitsdegradation of other components of the composition. In some embodiments,the transition metal complexing agent may be present in the compositionat a concentration of about 0.1 and 0.5 mg/mL, preferably in the rangeof about 0.1 and 0.4 mg/mL, more preferably in the range of about 0.1and 0.3 mg/mL, and most preferably about 0.2 mg/mL.

Preferably, transition metal complexing agents comprise one or more ofTPA (Tris[(2-pyridyl)methyl]amine), phanquinone(4,7-phenanthroline-5,6-dione), clioquinol (PN Gerolymatos SA),chloroquinol, penicillamine, trientine, N,N′-diethyldithiocarbamate(DDC), 2,3,2′-tetraamine (2,3,2′-tet), neocuproine,N,N,N′,N′-tetrakis(2-pyrdylmethyl) ethylenediamine (TPEN),1,10-phenanthroline (PHE), tetraethylenepentamine (TEPA), triethylenetetraamine and tris(2-carboxyethyl)phosphine (TCEP), bathophenanthrolinedisulfonic acid (BPADA), Disodium Edetate Dihydrate (EDTA), ethyleneglycol (bis) aminoethyl ether tetra acetic acid (EGTA), nitrilotriaceticacid, TIRON™, N,N-bis(2-hydroxyethyl)glycine (bicine),O,O′-bis(2-aminophenyl ethylene glycol)ethylenediamine-N,N,N′,N′-tetraacetic acid (BAPTA), trans-1,2-diaminocyclohexane-ethylenediamine-N,N,N′,N′-tetraacetic acid (CyDTA),1,3-diamino-2-hydroxy-propane-ethylenediamine-N,N,N′,N′-tetraacetic acid(DPTA-OH), ethylene-diamine-N,N′-dipropionic acid dihydrochloride(EDDP), ethylenediamine-N,N′-bis(methylenephosphonic acid) hemihydrate(EDDPO), ethylenediamine-N,N,N′,N′-tetrakis(methylenephosphonic acid)(EDTPO), N,N′-bis(2-hydroxybenzyl)ethylene diamine-N,N′-diacetic acid(HBED), 1,6-hexamethylenediamine-N,N,N′,N′-tetraacetic acid (HDTA, orHEDTA), N-(2-hydroxyethyl)iminodiacetic acid (HIDA), iminodiacetic acid(IDA), 1,2-diaminopropane-N,N,N′,N′-tetraacetic acid (methyl-EDTA),nitriltriacetic acid (NTA), nitrilotripropionic acid (NTP), nitrilotris(methylenephosphonic acid) trsodium salt (NTPO),triethylenetetramine-N,N,N′,N″,N″-hexaacetic acid (TTHA), bathocuproine,bathophenanthroline, TETA, citric acid, salicylic acid, and/or malicacid, including analogues, derivatives, and pharmaceutically acceptablesalts thereof. Preferably, the metal complexing agent comprises EDTA.

The present invention provides a composition that may include one ormore pH lowering agent. The term “pH lowering agent” refers to acomponent of a composition selected to lower the composition's pH. Insome embodiments, the pH lowering agent may be present in thecomposition such that the composition's pH is in a pharmaceuticallyacceptable range (i.e., not toxic or producing unacceptable sideeffects).

In some embodiments, the pH lowering agent may convert the active agentinto a salt that is soluble in the composition. In some embodiments, thepH lowering agent may convert epinephrine into a hydrochloride salt.

In some embodiments, the pH lowering agent may be present at aconcentration in the composition such that the composition's pH iswithin the limit on total acidity imposed by the USP 37-NF 32, S2,monograph. In some embodiments, the pH lowering agent may be present inthe composition at a concentration of in the range of about 0.001 to 1mg/mL, more preferably in the range of about 0.001 to 0.20 mg/mL, morepreferably in the range of about 0.001 to 0.10 mg/mL, more preferably inthe range of about 0.001 to 0.050 mg/mL and most preferably about 0.025mg/mL. In some embodiments, the pH lowering agent may be present in therange of about 0.1 to 1 mg/mL, preferably in the range of about 0.1 to0.5 mg/mL, and most preferably about 0.25 mg/mL.

Preferably, the pH lowering agent comprises one or more of acetic acid,adipic acid, ascorbic acid, citric acid, hydrochloric acid, lactic acid,malic acid, monopotassium phosphate, monosodium phosphate, phosphoricacid, pyrophosphoric acid, succinic acid, sulfuric acid, and or tartaricacid. Preferably, the pH lowering agent comprises hydrochloric acid. Incertain embodiments, the pH lowering agent may be a portion of thebuffer system in conjugation with a pH raising agent.

The present invention provides for a composition that may comprise atleast one tonicity regulating agent. The term “tonicity” refers to theeffective osmotic pressure equivalent of a solution or composition. Insome embodiments, the tonicity regulating agent may be present in thecomposition to maintain the tonicity of the composition in aphysiological acceptable range. In some embodiments, the tonicityregulating agent may be an osmotic pressure regulating agent.

Preferably, the tonicity regulating agent comprises one or more ofglucose, glycerin, hydroxypropyl methyl cellulose, mannitol,polysorbate, propylene glycol, sodium bromide, sodium chloride, sodiumiodide, sorbitol, urea, xylitol, and/or combinations thereof.Preferably, the tonicity regulating agent comprises sodium chloride.

In some embodiments, the tonicity regulating agent may be present in thecomposition at a concentration of in the range of about 4 and 9 mg/mL,preferably in the range of about 5 and 8 mg/mL, and most preferably inthe range of about 6 and 7.5 mg/mL. In some embodiments, the tonicityregulating agent may be present at a concentration of about 7.3 mg/mL.In some embodiments, the tonicity regulating agent may be present at aconcentration of about 6.15 mg/mL.

The present invention provides for a composition that may optionallycomprise one or more preservatives. The term “preservative” refers to asubstance present in a composition which can, at least in part, preventand/or reduce decomposition of the composition. In some embodiments, thepreservative may prevent and/or reduce decomposition of the compositionby microbial growth in the composition. Preferably, the preservative ispharmaceutically acceptable. In some embodiments, the composition isfree of preservatives.

In some embodiments, the preservative may be present in the compositionat a concentration that allows for a multi-dose formulation of thecomposition. In some embodiments, the preservative may be present in thecomposition at a concentration that prevents and/or reducesdecomposition of unused portions of the composition in a multi-doseformulation. In some embodiments, the preservative may allow for up toabout 14 days of use, preferably up to about 30 days of use, morepreferably up to about 60 days of use, and most preferably up to about90 days of use of a multi-dose formulation of the composition.

In some embodiments, the preservative may be present in the compositionat a concentration of in the range of about 1 to 10 mg/mL, preferably inthe range of about 3 and 7 mg/mL, more preferably in the range of about4 and 6 mg/mL, more preferably in the range of about 4.5 and 5.5 mg/mL,and most preferably at about 5 mg/mL.

Preferably, preservatives comprise one or more of benzalkonium chloride,benzethonium chloride, benzoic acid, benzyl alcohol, benzyl paraben,bronopol, butyl paraben, cetrimide, cetylpyridinium chloride,chlorbutanol, chlorhexidine, chlorocresol, chloroxylenol, cresol, ethylalcohol, ethyl paraben, ethylparaben, glycerin, hexetidine, imidurea,isobutyl paraben, meta-cresol, methyl paraben, methylparaben, phenol,phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate,p-hydroxybenzoic acid esters, polyhexamethylene biguanide (“PHMB”),potassium sorbate, propyl paraben, propylene glycol, sodium benzoate,sodium perborate, sodium propionate, sorbic acid, stabilized thimerosal,and/or thimerosal. Preferably, the preservative comprises chlorobutanol.

The present invention provides for a composition that can optionallycomprise one or more solvents. In some embodiments, the solvent may beacceptable for pharmaceutical administration. Examples of methods ofpharmaceutical administration include, but are not limited to,subcutaneous, intracameral, intravenous, and intramuscular injection,infusion, intra-arterial administration, intracardiac injection,endotracheal administration, intraosseous administration, oralinhalation, topical administration, and as ophthalmic irrigation.

Preferably, the solvent comprises one or more of acetic acid, acetone,acetonitrile, animal oil, aqueous buffer, benzene, bisabolol, butanol,carbon tetrachloride, chlorobenzene, chloroform, dimethylformamide,dioxane, essential oil, ethanol, ethyl acetate, ethyl oleate, ethylenechloride, fatty acid esters, glycerin, glycofurol, hexane, hydrogenatedvegetable oil, isopropanol, isopropyl alcohol, isopropyl myristate,isopropyl palmitate, methanol, methylene chloride, mineral oil,polyethylene glycol, polyol, propylene glycol, silicone fluid glyceride,squalane, terpene, tetrahydrofuran, toluene, triacetin, tributylcitrate, triethyl citrate, vegetable oil, and/or water. Preferably thesolvent comprises water.

In some embodiments, the solvent may be present in an amount that bringsthe composition to a final volume which is suitable for administration.In some embodiments, the final volume may be 1 mL. In some embodiments,the final volume may be 30 mL.

The present invention provides for a composition that may have a lowlevel of impurities. The term “impurity” refers to an undesiredsubstance in a composition. In some embodiments, an amount of impuritiesmay be present in an initial composition and/or may be formed after acertain period of shelf life of a composition. In some embodiments,impurities may be formed via degradation of one or more components ofthe composition. Sources of degradation include, but are not limited to,oxidation, racemization, sulfite addition, visible light, ultravioletlight, moisture, heat, changes in pH, and composition componentinteractions. Unless indicated otherwise, the percentages of impuritiesexpressed herein are expressed as % w/w of the active agent.

In some embodiments, the composition may have no more than about 20% oftotal impurities after a certain period of shelf life, more preferablyno more than about 19.5%, more preferably no more than about 19%, morepreferably no more than about 18.5%, preferably no more than about 18%,more preferably no more than about 17.5%, more preferably no more thanabout 17%, more preferably no more than about 16.5%, more preferably nomore than about 16%, more preferably no more than about 15.5%, morepreferably no more than about 15%, more preferably no more than about14.5%, more preferably no more than about 14%, more preferably no morethan about 13.5%, more preferably no more than about 13%, morepreferably no more than about 12.5%, more preferably no more than about12%, more preferably no more than about 11.5%, more preferably no morethan about 11%, more preferably no more than about 10.5%, morepreferably no more than about 10%, more preferably no more than about9.5%, more preferably no more than about 9%, more preferably no morethan about 8.5%, more preferably no more than about 8%, more preferablyno more than about 7.5%, more preferably no more than about 7%, morepreferably no more than about 6.5%, more preferably no more than about6%, more preferably no more than about 5.5%, and most preferably no morethan about 5%.

In some embodiments, the composition may have no more than about 20% oftotal impurities after a certain period of shelf life, preferably nomore than about 19.9%, more preferably no more than about 19.8%, morepreferably no more than about 19.7%, more preferably no more than about19.6%, more preferably no more than about 19.5%, more preferably no morethan about 19.4%, more preferably no more than about 19.3%, morepreferably no more than about 19.2%, more preferably no more than about19.1%, more preferably no more than about 19%, more preferably no morethan about 18.9%, more preferably no more than about 18.8%, morepreferably no more than about 18.7%, more preferably no more than about18.6%, more preferably no more than about 18.5%, more preferably no morethan about 18.4%, more preferably no more than about 18.3%, morepreferably no more than about 18.2%, more preferably no more than about18.1%, more preferably no more than about 18%, more preferably no morethan about 17.9%, more preferably no more than about 17.8%, morepreferably no more than about 17.7%, more preferably no more than about17.6%, more preferably no more than about 17.5%, more preferably no morethan about 17.4%, more preferably no more than about 17.3%, morepreferably no more than about 17.2%, more preferably no more than about17.1%, more preferably no more than about 17%, more preferably no morethan about 16.9%, more preferably no more than about 16.8%, morepreferably no more than about 16.7%, more preferably no more than about16.6%, more preferably no more than about 16.5%, more preferably no morethan about 16.4%, more preferably no more than about 16.3%, morepreferably no more than about 16.2%, more preferably no more than about16.1%, more preferably no more than about 16%, more preferably no morethan about 15.9%, more preferably no more than about 15.8%, morepreferably no more than about 15.7%, more preferably no more than about15.6%, more preferably no more than about 15.5%, more preferably no morethan about 15.4%, more preferably no more than about 15.3%, morepreferably no more than about 15.2%, more preferably no more than about15.1%, more preferably no more than about 15%, more preferably no morethan about 14.9%, more preferably no more than about 14.8%, morepreferably no more than about 14.7%, more preferably no more than about14.6%, more preferably no more than about 14.5%, more preferably no morethan about 14.4%, more preferably no more than about 14.3%, morepreferably no more than about 14.2%, more preferably no more than about14.1%, more preferably no more than about 14%, more preferably no morethan about 13.9%, more preferably no more than about 13.8%, morepreferably no more than about 13.7%, more preferably no more than about13.6%, more preferably no more than about 13.5%, more preferably no morethan about 13.4%, more preferably no more than about 13.3%, morepreferably no more than about 13.2%, more preferably no more than about13.1%, more preferably no more than about 13%, more preferably no morethan about 12.9%, more preferably no more than about 12.8%, morepreferably no more than about 12.7%, more preferably no more than about12.6%, more preferably no more than about 12.5%, more preferably no morethan about 12.4%, more preferably no more than about 12.3%, morepreferably no more than about 12.2%, more preferably no more than about12.1%, more preferably no more than about 12%, more preferably no morethan about 11.9%, more preferably no more than about 11.8%, morepreferably no more than about 11.7%, more preferably no more than about11.6%, more preferably no more than about 11.5%, more preferably no morethan about 11.4%, more preferably no more than about 11.3%, morepreferably no more than about 11.2%, more preferably no more than about11.1%, more preferably no more than about 11%, more preferably no morethan about 10.9%, more preferably no more than about 10.8%, morepreferably no more than about 10.7%, more preferably no more than about10.6%, more preferably no more than about 10.5%, more preferably no morethan about 10.4%, more preferably no more than about 10.3%, morepreferably no more than about 10.2%, more preferably no more than about10.1%, more preferably no more than about 10%, more preferably no morethan about 9.9%, more preferably no more than about 9.8%, morepreferably no more than about 9.7%, more preferably no more than about9.6%, more preferably no more than about 9.5%, more preferably no morethan about 9.4%, more preferably no more than about 9.3%, morepreferably no more than about 9.2%, more preferably no more than about9.1%, more preferably no more than about 9%, more preferably no morethan about 8.9%, more preferably no more than about 8.8%, morepreferably no more than about 8.7%, more preferably no more than about8.6%, more preferably no more than about 8.5%, more preferably no morethan about 8.4%, more preferably no more than about 8.3%, morepreferably no more than about 8.2%, more preferably no more than about8.1%, more preferably no more than about 8%, more preferably no morethan about 7.9%, more preferably no more than about 7.8%, morepreferably no more than about 7.7%, more preferably no more than about7.6%, more preferably no more than about 7.5%, more preferably no morethan about 7.4%, more preferably no more than about 7.3%, morepreferably no more than about 7.2%, more preferably no more than about7.1%, more preferably no more than about 7%, more preferably no morethan about 6.9%, more preferably no more than about 6.8%, morepreferably no more than about 6.7%, more preferably no more than about6.6%, more preferably no more than about 6.5%, more preferably no morethan about 6.4%, more preferably no more than about 6.3%, morepreferably no more than about 6.2%, more preferably no more than about6.1%, more preferably no more than about 6%, more preferably no morethan about 5.9%, more preferably no more than about 5.8%, morepreferably no more than about 5.7%, more preferably no more than about5.6%, more preferably no more than about 5.5%, more preferably no morethan about 5.4%, more preferably no more than about 5.3%, morepreferably no more than about 5.2%, more preferably no more than about5.1%, more preferably no more than about 5%, more preferably no morethan about 4.9%, more preferably no more than about 4.8%, morepreferably no more than about 4.7%, more preferably no more than about4.6%, more preferably no more than about 4.5%, more preferably no morethan about 4.4%, more preferably no more than about 4.3%, morepreferably no more than about 4.2%, more preferably no more than about4.1%, more preferably no more than about 4%, more preferably no morethan about 3.9%, more preferably no more than about 3.8%, morepreferably no more than about 3.7%, more preferably no more than about3.6%, more preferably no more than about 3.5%, more preferably no morethan about 3.4%, more preferably no more than about 3.3%, morepreferably no more than about 3.2%, more preferably no more than about3.1%, more preferably no more than about 3%, more preferably no morethan about 2.9%, more preferably no more than about 2.8%, morepreferably no more than about 2.7%, more preferably no more than about2.6%, more preferably no more than about 2.5%, more preferably no morethan about 2.4%, more preferably no more than about 2.3%, morepreferably no more than about 2.2%, more preferably no more than about2.1%, more preferably no more than about 2%, more preferably no morethan about 1.9%, more preferably no more than about 1.8%, morepreferably no more than about 1.7%, more preferably no more than about1.6%, more preferably no more than about 1.5%, more preferably no morethan about 1.4%, more preferably no more than about 1.3%, morepreferably no more than about 1.2%, more preferably no more than about1.1%, more preferably no more than about 1%, more preferably no morethan about 0.9%, more preferably no more than about 0.8%, morepreferably no more than about 0.7%, more preferably no more than about0.6%, more preferably no more than about 0.5%, more preferably no morethan about 0.4%, more preferably no more than about 0.3%, morepreferably no more than about 0.2%, more preferably no more than about0.1%, and most preferably about 0%.

In some embodiments, the concentration of impurities present in thecomposition after a certain period of shelf life may be attributed atleast partially to degradation of components of the composition. In someembodiments, the concentration of impurities present in the compositionat the end of shelf life may be attributed at least partially toepinephrine degradation. In some embodiments, epinephrine degradationmay be a result of physical or chemical stress. Examples of stressesinclude, but are not limited to, oxygen, pH, bisulfite, light, processsurface compatibility, and soluble trace metals.

In some embodiments, components of the composition may be present at aconcentration that lowers the amount of impurities after a certainperiod of shelf life that would otherwise be present in the absence ofthe components. In some embodiments, components of the composition maybe present at a concentration that at least partially inhibits formationof impurities in the composition.

The present invention provides for a composition that may have a lowlevel of D-epinephrine.

In some embodiments, the concentration of D-epinephrine in thecomposition after a certain period of shelf life may be no more thanabout 9.5%, preferably no more than about 9%, more preferably no morethan about 8.5%, more preferably no more than about 8%, more preferablyno more than about 7.5%, more preferably no more than about 7%, morepreferably no more than about 6.5%, more preferably no more than about6%, more preferably no more than about 5.5%, more preferably no morethan about 5%, more preferably no more than about 4.5%, more preferablyno more than about 4%, more preferably no more than about 3.5%, morepreferably no more than than about 3%, more preferably no more thanabout 2.5%, more preferably no more than about 2%, more preferably nomore than about 1.5%, more preferably no more than about 1%, and mostpreferably no more than about 0.5%.

In some embodiments, the concentration of D-epinephrine in thecomposition after a period of shelf life may be no more than about 9.5%,preferably no more than about 9.4%, more preferably no more than about9.3%, more preferably no more than about 9.2%, more preferably no morethan about 9.1%, more preferably no more than about 9%, more preferablyno more than about 8.9%, more preferably no more than about 8.8%, morepreferably no more than about 8.7%, more preferably no more than about8.6%, more preferably no more than about 8.5%, more preferably no morethan about 8.4%, more preferably no more than about 8.3%, morepreferably no more than about 8.2%, more preferably no more than about8.1%, more preferably no more than about 8%, more preferably no morethan about 7.9%, more preferably no more than about 7.8%, morepreferably no more than about 7.7%, more preferably no more than about7.6%, more preferably no more than about 7.5%, more preferably no morethan about 7.4%, more preferably no more than about 7.3%, morepreferably no more than about 7.2%, more preferably no more than about7.1%, more preferably no more than about 7%, more preferably no morethan about 6.9%, more preferably no more than about 6.8%, morepreferably no more than about 6.7%, more preferably no more than about6.6%, more preferably no more than about 6.5%, more preferably no morethan about 6.4%, more preferably no more than about 6.3%, morepreferably no more than about 6.2%, more preferably no more than about6.1%, more preferably no more than about 6%, more preferably no morethan about 5.9%, more preferably no more than about 5.8%, morepreferably no more than about 5.7%, more preferably no more than about5.6%, more preferably no more than about 5.5%, more preferably no morethan about 5.4%, more preferably no more than about 5.3%, morepreferably no more than about 5.2%, more preferably no more than about5.1%, more preferably no more than about 5%, more preferably no morethan about 4.9%, more preferably no more than about 4.8%, morepreferably no more than about 4.7%, more preferably no more than about4.6%, more preferably no more than about 4.5%, more preferably no morethan about 4.4%, more preferably no more than about 4.3%, morepreferably no more than about 4.2%, more preferably no more than about4.1%, more preferably no more than about 4%, more preferably no morethan about 3.9%, more preferably no more than about 3.8%, morepreferably no more than about 3.7%, more preferably no more than about3.6%, more preferably no more than about 3.5%, more preferably no morethan about 3.4%, more preferably no more than about 3.3%, morepreferably no more than about 3.2%, more preferably no more than about3.1%, more preferably no more than about 3%, more preferably no morethan about 2.9%, more preferably no more than about 2.8%, morepreferably no more than about 2.7%, more preferably no more than about2.6%, more preferably no more than about 2.5%, more preferably no morethan about 2.4%, more preferably no more than about 2.3%, morepreferably no more than about 2.2%, more preferably no more than about2.1%, more preferably no more than about 2%, more preferably no morethan about 1.9%, more preferably no more than about 1.8%, morepreferably no more than about 1.7%, more preferably no more than about1.6%, more preferably no more than about 1.5%, more preferably no morethan about 1.4%, more preferably no more than about 1.3%, morepreferably no more than about 1.2%, more preferably no more than about1.1%, more preferably no more than about 1%, more preferably no morethan about 0.9%, more preferably no more than about 0.8%, morepreferably no more than about 0.7%, more preferably no more than about0.6%, more preferably no more than about 0.5%, more preferably no morethan about 0.4%, more preferably no more than about 0.3%, morepreferably no more than about 0.2%, more preferably no more than about0.1%, more preferably no more than about 0.09%, more preferably no morethan about 0.08%, more preferably no more than about 0.07%, morepreferably no more than about 0.06%, more preferably no more than about0.05%, more preferably no more than about 0.04%, more preferably no morethan about 0.03%, more preferably no more than about 0.02%, and mostpreferably no more than about 0.01%.

In some embodiments, the initial concentration of D-epinephrine in thecomposition may be no more than about 2.5%, preferably no more thanabout 2%, more preferably no more than about 1.5%, more preferably nomore than about 1%, and most preferably no more than about 0.5%.

In some embodiments, components of the composition may be present atconcentrations that reduce the formation of D-epinephrine that wouldotherwise occur in the absence of the components. In some embodiments,components of the composition may be present at concentrations thatreduce D-epinephrine formation by at least about 15%, more preferably atleast about 33%, and most preferably at least about 50%.

In some embodiments, components of the composition may be present at aconcentration that reduces the rate and/or extent of D-epinephrineformation after a certain period of shelf life. In some embodiments,components of the composition may reduce D-epinephrine formation byraising the initial pH of the composition and/or by increasing the pHrange of the composition over shelf life. In some embodiments, the pH ofthe composition may be raised at least partially by the pH raisingagent. Preferably, the raised pH of the composition does not result inlarge increases of other degradants of epinephrine.

The present invention provides for a composition that may have a lowlevel of epinephrine sulfonic acid (ESA). In some embodiments, theconcentration of ESA in the composition after a certain period of shelflife may be no more than about 14.5%, preferably no more than about 14%,more preferably no more than about 13.5%, more preferably no more thanabout 13%, more preferably no more than about 12.5%, more preferably nomore than about 12%, more preferably no more than about 11.5%, morepreferably no more than about 11%, more preferably no more than about10.5%, more preferably no more than about 10%, more preferably no morethan about 9.5%, more preferably no more than about 9%, more preferablyno more than about 8.5%, more preferably no more than about 8%, morepreferably no more than about 7.5%, more preferably no more than about7%, more preferably no more than about 6.5%, more preferably no morethan about 6%, and most preferably no more than about 5.5%.

In some embodiments, the concentration of ESA in the composition after acertain period of shelf life may be no more than about 14.5%, preferablyno more than about 14.4%, more preferably no more than about 14.3%, morepreferably no more than about 14.2%, more preferably no more than about14.1%, more preferably no more than about 14%, more preferably no morethan about 13.9%, more preferably no more than about 13.8%, morepreferably no more than about 13.7%, more preferably no more than about13.6%, more preferably no more than about 13.5%, more preferably no morethan about 13.4%, more preferably no more than about 13.3%, morepreferably no more than about 13.2%, more preferably no more than about13.1%, more preferably no more than about 13%, more preferably no morethan about 12.9%, more preferably no more than about 12.8%, morepreferably no more than about 12.7%, more preferably no more than about12.6%, more preferably no more than about 12.5%, more preferably no morethan about 12.4%, more preferably no more than about 12.3%, morepreferably no more than about 12.2%, more preferably no more than about12.1%, more preferably no more than about 12%, more preferably no morethan about 11.9%, more preferably no more than about 11.8%, morepreferably no more than about 11.7%, more preferably no more than about11.6%, more preferably no more than about 11.5%, more preferably no morethan about 11.4%, more preferably no more than about 11.3%, morepreferably no more than about 11.2%, more preferably no more than about11.1%, more preferably no more than about 11%, more preferably no morethan about 10.9%, more preferably no more than about 10.8%, morepreferably no more than about 10.7%, more preferably no more than about10.6%, more preferably no more than about 10.5%, more preferably no morethan about 10.4%, more preferably no more than about 10.3%, morepreferably no more than about 10.2%, more preferably no more than about10.1%, more preferably no more than about 10%, more preferably no morethan about 9.9%, more preferably no more than about 9.8%, morepreferably no more than about 9.7%, more preferably no more than about9.6%, more preferably no more than about 9.5%, more preferably no morethan about 9.4%, more preferably no more than about 9.3%, morepreferably no more than about 9.2%, more preferably no more than about9.1%, more preferably no more than about 9%, more preferably no morethan about 8.9%, more preferably no more than about 8.8%, morepreferably no more than about 8.7%, more preferably no more than about8.6%, more preferably no more than about 8.5%, more preferably no morethan about 8.4%, more preferably no more than about 8.3%, morepreferably no more than about 8.2%, more preferably no more than about8.1%, more preferably no more than about 8%, more preferably no morethan about 7.9%, more preferably no more than about 7.8%, morepreferably no more than about 7.7%, more preferably no more than about7.6%, more preferably no more than about 7.5%, more preferably no morethan about 7.4%, more preferably no more than about 7.3%, morepreferably no more than about 7.2%, more preferably no more than about7.1%, more preferably no more than about 7%, more preferably no morethan about 6.9%, more preferably no more than about 6.8%, morepreferably no more than about 6.7%, more preferably no more than about6.6%, more preferably no more than about 6.5%, more preferably no morethan about 6.4%, more preferably no more than about 6.3%, morepreferably no more than about 6.2%, more preferably no more than about6.1%, more preferably no more than about 6%, more preferably no morethan about 5.9%, more preferably no more than about 5.8%, morepreferably no more than about 5.7%, more preferably no more than about5.6%, more preferably no more than about 5.5%, more preferably no morethan about 5.4%, more preferably no more than about 5.3%, morepreferably no more than about 5.2%, more preferably no more than about5.1%, more preferably no more than about 5.0%, more preferably no morethan about 4.9%, more preferably no more than about 4.8%, morepreferably no more than about 4.7%, more preferably no more than about4.6%, more preferably no more than about 4.5%, more preferably no morethan about 4.4%, more preferably no more than about 4.3%, morepreferably no more than about 4.2%, more preferably no more than about4.1%, more preferably no more than about 4.0%, more preferably no morethan about 3.9%, more preferably no more than about 3.8%, morepreferably no more than about 3.7%, more preferably no more than about3.6%, more preferably no more than about 3.5%, more preferably no morethan about 3.4%, more preferably no more than about 3.3%, morepreferably no more than about 3.2%, more preferably no more than about3.1%, more preferably no more than about 3.0%, more preferably no morethan about 2.9%, more preferably no more than about 2.8%, morepreferably no more than about 2.7%, more preferably no more than about2.6%, more preferably no more than about 2.5%, more preferably no morethan about 2.4%, more preferably no more than about 2.3%, morepreferably no more than about 2.2%, more preferably no more than about2.1%, more preferably no more than about 2.0%, more preferably no morethan about 1.9%, more preferably no more than about 1.8%, morepreferably no more than about 1.7%, more preferably no more than about1.6%, more preferably no more than about 1.5%, more preferably no morethan about 1.4%, more preferably no more than about 1.3%, morepreferably no more than about 1.2%, more preferably no more than about1.1%, more preferably no more than about 1.0%, more preferably no morethan about 0.9%, more preferably no more than about 0.8%, morepreferably no more than about 0.7%, more preferably no more than about0.6%, more preferably no more than about 0.5%, more preferably no morethan about 0.4%, more preferably no more than about 0.3%, morepreferably no more than about 0.2%, more preferably no more than about0.1%, and most preferably about 0%.

In some embodiments, the initial concentration of ESA in the compositionmay be no more than about 0.5%, preferably no more than about 0.2%, morepreferably no more than about 0.1%, more preferably no more than about0.05%, and most preferably no more than about 0.025%.

In some embodiments, ESA formation may be influenced by anoxygen-dependent mechanism such that reducing oxygen accessible to thecomposition may reduce the rate of ESA formation. In some embodiments,oxygen present in the headspace of a vial containing the composition maybe reduced by replacing some or all of the oxygen with a gas other thanoxygen, preferably an inert gas, thereby at least partially reducing therate of ESA production. In some embodiments, some or all of the oxygenpresent in the headspace of a vial containing the composition may bereplaced with nitrogen and/or argon.

In some embodiments, the headspace gas of the composition is manipulatedsuch that degradation of components in the composition is minimized. Theterm “headspace” refers to the gas space above the product in acontainer. The headspace gas may be manipulated in order to reduce theamount of oxygen present therein. In some embodiments, the amount ofoxygen in the headspace gas is no more than about 15% v/v of the totalgas, preferably no more than about 13%, more preferably no more thanabout 10%, more preferably no more than about 5%, more preferably nomore than about 2.5%, more preferably no more than about 1%, and mostpreferably about 0%.

In some embodiments of the present invention, the concentrations ofcertain components of the composition may be reduced such that theconcentration of ESA in the composition after a period of shelf life isreduced. In some embodiments, the concentration of ESA present in thecomposition after a period of shelf life is reduced at least partiallyby reducing the initial concentration of antioxidant in the composition.In some embodiments, the concentrations of certain components of thecomposition, such as initial concentration of antioxidant, are reducedsuch that the rate of ESA production is reduced by at least about 25%,more preferably at least about 50%, and most preferably at least about75% after a certain period of shelf life, as compared to a compositionin which the certain components are not present.

The present invention provides for a composition that may have a lowlevel of oxidative degradants. The term “oxidative degradant” refers toany impurity that may be at least partially attributed to an oxidationreaction involving one or more components of the composition. In someembodiments, the oxidative degradants may be formed via oxidation ofepinephrine. Examples of oxidative degradants include, but are notlimited to, adrenalone, adrenochrome sulfonate, adrenochrome,adrenolutin, melanins, and analogs thereof.

In some embodiments, the concentration of one of the oxidativedegradants in the composition after a certain period of shelf life maybe no more than about 1%, preferably no more than about 0.9%, morepreferably no more than about 0.8%, more preferably no more than about0.7%, more preferably no more than about 0.6%, more preferably no morethan about 0.5%, more preferably no more than about 0.4%, morepreferably no more than about 0.3%, more preferably no more than about0.2%, more preferably no more than about 0.1%, more preferably no morethan about 0.05%, more preferably no more than about 0.04%, morepreferably no more than about 0.03%, more preferably no more than about0.02%, and most preferably no more than about 0.01%, after a certainperiod of shelf life.

In some embodiments, the concentration of more than one or all of theoxidative degradants in the composition after a certain period of shelflife may be no more than about 1%, preferably no more than about 0.9%,more preferably no more than about 0.8%, more preferably no more thanabout 0.7%, more preferably no more than about 0.6%, more preferably nomore than about 0.5%, more preferably no more than about 0.4%, morepreferably no more than about 0.3%, more preferably no more than about0.2%, more preferably no more than about 0.1%, more preferably no morethan about 0.05%, more preferably no more than about 0.04%, morepreferably no more than about 0.03%, more preferably no more than about0.02%, and most preferably no more than about 0.01%, after a certainperiod of shelf life.

In some embodiments, the concentration of oxidative degradants presentin the composition after a certain period of shelf life may be such thatthe composition does not undergo a physical change. Examples of physicalchange include, but are not limited to, color change and insolubleparticle formation.

In some embodiments, the rate of formation and/or the concentration ofoxidative degradants in the composition may be reduced after a certainperiod of shelf life by other components of the composition. In someembodiments, the rate of formation and/or the concentration of oxidativedegradants in the composition may be reduced by the antioxidant. In someembodiments, the rate of formation and/or the concentration of oxidativedegradants in the composition may be reduced by reducing the amount ofoxygen accessible to the composition. In some embodiments, the rate offormation and/or the concentration of oxidative degradants in thecomposition may be reduced by replacing some or all of the oxygen in theheadspace of a vial containing the composition with non-oxygen gas,preferably an inert gas such as nitrogen and/or argon.

In some embodiments, the present invention may have low levels ofdegradants Impurity A, Impurity B, and/or Unknown C.

In some embodiments, Impurity A may be a compound with the followingstructure:

In some embodiments, Impurity B may be a compound with the followingstructure:

In some embodiments, Unknown C may be characterized by a λ_(max) ofabout 380 nm.

In some embodiments, Unknown C may be characterized by its elution peakusing HILIC (Hydrophilic Interaction Liquid Chromatography), whichdiffers from one substance to another.

In some embodiments, Unknown C may be characterized using an isocraticHILIC separation of mixtures according to the conditions described inTables 1-3.

Table 1 lists the compositions of the mobile phases that may be used forisocratic HILIC separations.

TABLE 1 Composition of Isocratic Mobile Phases Mobile Phase A 100 mLAmmonium Formate Buffer 900 mL Acetonitrile Mobile Phase B 100 mLAmmonium Formate Buffer 400 mL Water 500 mL Acetonitrile

The Ammonium Formate Buffer of Table 1 may be prepared as follows: 1.5 gof Sodium Chloride, 5.0 mL of Formic Acid, and 3.0 mL of 6N AmmoniumHydroxide is added to 1.0 L of water. The solution may be vacuumfiltered through a 0.45 μm nylon membrane. The preparation may be scaledup where necessary.

Table 2 lists different conditions that may be used for isocratic HILICseparations.

TABLE 2 Isocratic Mobile Phases for HILIC Separation Isocratic MobileMobile Water % Phase A % Phase B % 15 87.5 12.5 18 80 20 22 70 30

An Epinephrine HILIC Impurities Marker Solution (EpiHILIC IMS) may beprepared by first preparing an Epinephrine/Degradant Bulk Solution asfollows: In a clean, clear, 2 L glass bottle, dissolving sodium chloride(6.147 g) and tartaric acid (2.251 g) in about 800 mL of water, addingsodium hydroxide 5.0N (6.25 mL) to the solution and mixing, addingsodium metabisulfite (1.425 g) to the solution and mixing to dissolve,adding epinephrine (1.1334 g) as a solution in HCl 1.0N (6.6 mL) to thesolution, and then adding water to a final solution weight of 1005.9,and mixing thoroughly. The Epinephrine/Degradant Bulk Solution may thenbe allowed to sit in a capped bottle in the hood, under fluorescentlight with a light exposure at about 1000 lux, for 12 days with lightscontinuously on.

The Epinephne HILIC Impurities MarkerSolution (EpiHILIC IMS) may then beprepared in a 500 mL clearglass bottle as follows: addingEpinephlne/Degradant Bulk Solution (500 mL), adding HCl 1.0 N (6.0 ml)and then mixing (with a final pH at 3.2), adding ESA (9.5 mg) and mixingto dissolve, adding adrenochrome (5.6 mg) and mixing to dissolve,preparing adrenalone stock mixture by withdrawing 10 mL of the solutionthen adding and dissolving adrenalone HCl (4.7 mg), adding adrenalonestock mixture (0.8 ml) back into the bulk mixture, and mixingthoroughly.

An Epinephrine HILIC Impurities Marker Solution (EpiHILIC IMS) whichcomprises Unknown C may be analyzed using the conditions listed in Table3.

TABLE 3 Chromatographic Conditions for Isocratic HILIC Separation ColumnSeQuant ZIC HILIC PEEK, 150 × 4.6 mm, 3.5 μm particle size, 100 Å poresize Flow Rate 1 mL/min Injection Volume 25 μL Column Temperature 35° C.Sample preparation Dilute exactly 1 volume of sample (nominal strengthof 1 mg/mL epinephrine) with exactly 3 volumes of acetonitrile. DetectorSignal-280 nm, Bandwidth-10 nm; Reference-Off Signal-346 nm,Bandwidth-10 nm; Reference-Off Signal-380 nm, Bandwidth-10 nm;Reference-Off Signal-292 nm, Bandwidth-10 nm; Reference-Off Run Time 35minutes Data Acquisition Integrate impurities between 0 and 30 minutes

Tables 4 lists the relevant characteristic peaks of isocratic HILICseparation according to the above specifications at a detectionwavelength of 346 nm using the 15% isocratic water preparation accordingto Table 2.

TABLE 4 Chromatography of the Impurities Marker Solution using isocraticHILIC with 15% water and 346 nM Detection Time (Minutes) Absorbance (AU)Unknown C approx. 21 approx. 0.001

Tables 5 lists the relevant characteristic peaks of isocratic HILICseparation according to the above specifications at a detectionwavelength of 346 nm using the 18% isocratic water preparation accordingto Table 2.

TABLE 5 Chromatography of the Impurities Marker Solution using isocraticHILIC with 18% water and 346 nM Detection Time (Minutes) Absorbance (AU)Unknown C approx. 14 approx. 0.002

Table 6 lists the relevant characteristic peaks of isocratic HILICseparation according to the above specifications at a detectionwavelength of 346 nm using the 22% isocratic water preparation accordingto Table 2.

TABLE 6 Chromatography of the Impurities Marker Solution using isocraticHILIC with 22% water and 346 nM Detection Time (Minutes) Absorbance (AU)Unknown C approx. 8.5 approx. 0.002

The chromatography outputs of the Impurities Marker Solution usingisocratic HILIC as described is also shown in FIG. 1 (15% water), FIG. 2(18% water), and FIG. 3 (22% water).

In some embodiments, Unknown C may be characterized using a gradientHILIC method. An Epinephrine HILIC Impurities Marker Solution (EpiHILICIMS) which comprises Unknown C may be analyzed using the conditionslisted in Table 7 and 8.

Table 7 lists the compositions of the mobile phases that may be used forgradient HILIC separations.

TABLE 7 Composition of Gradient Mobile Phases Mobile Phase A 100 mLAmmonium Formate Buffer 900 mL Acetonitrile Mobile Phase A 100 mLAmmonium Formate Buffer 400 mL Water 500 mL Acetonitrile

Table 8 lists the solvent program for the gradient HILIC method.

TABLE 8 Solvent program for the gradient HILICmethod. Mobile PhaseComposition Time (min) Flow (mL/min) Water % A % B % Initial 1.0 15.2 8713 12 1.0 15.2 87 13 22 1.0 22 70 30 25 1.0 22 70 30 28 1.0 15.2 87 1335 1.0 15.2 87 13

The chromatographic conditions for the gradient HILIC are the same asthose listed in Table 3.

The chromatography outputs of the Impurities Marker Solution usinggradient HILIC according to the above specifications at detectionwavelengths of 280 nm, 292 nm, 346 nm, and 380 nm are shown in FIG. 4 .

In some embodiments, the concentration Impurity A, Impurity B, andUnknown C together in the composition after a certain period of shelflife is no more than about 5%, preferably no more than about 4%, morepreferably no more than about 3%, more preferably no more than about 2%,more preferably no more than about 1%, and most preferably no more thanabout 0.5%.

In some embodiments, the concentration of Impurity A in the compositionafter a certain period of shelf life may be no more than about 2%,preferably no more than about 1.9%, more preferably no more than about1.8%, more preferably no more than about 1.7%, more preferably no morethan about 1.6%, more preferably no more than about 1.5%, morepreferably no more than about 1.4%, more preferably no more than about1.3%, more preferably no more than about 1.2%, more preferably no morethan about 1.1%, more preferably no more than about 1%, more preferablyno more than about 0.9%, more preferably no more than about 0.8%, morepreferably no more than about 0.7%, more preferably no more than about0.6%, more preferably no more than about 0.5%, more preferably no morethan about 0.4%, more preferably no more than about 0.3%, morepreferably no more than about 0.2%, and most preferably no more thanabout 0.1%.

In some embodiments, the concentration of Impurity B in the compositionafter a certain period of shelf life may be no more than about 2%,preferably no more than about 1.9%, more preferably no more than about1.8%, more preferably no more than about 1.7%, more preferably no morethan about 1.6%, more preferably no more than about 1.5%, morepreferably no more than about 1.4%, more preferably no more than about1.3%, more preferably no more than about 1.2%, more preferably no morethan about 1.1%, more preferably no more than about 1%, more preferablyno more than about 0.9%, more preferably no more than about 0.8%, morepreferably no more than about 0.7%, more preferably no more than about0.6%, more preferably no more than about 0.5%, more preferably no morethan about 0.4%, more preferably no more than about 0.3%, morepreferably no more than about 0.2%, and most preferably no more thanabout 0.1%.

In some embodiments, the concentration of Unknown C in the compositionafter a certain period of shelf life may be no more than about 2%,preferably no more than about 1.9%, more preferably no more than about1.8%, more preferably no more than about 1.7%, more preferably no morethan about 1.6%, more preferably no more than about 1.5%, morepreferably no more than about 1.4%, more preferably no more than about1.3%, more preferably no more than about 1.2%, more preferably no morethan about 1.1%, more preferably no more than about 1%, more preferablyno more than about 0.9%, more preferably no more than about 0.8%, morepreferably no more than about 0.7%, more preferably no more than about0.6%, more preferably no more than about 0.5%, more preferably no morethan about 0.4%, more preferably no more than about 0.3%, morepreferably no more than about 0.2%, and most preferably no more thanabout 0.1%.

In some embodiments, the presence of Impurity A, Impurity B, and/orUnknown C in the composition may be at least partially related to thepresence of oxygen such that reducing oxygen accessible to thecomposition may reduce the amount of Impurity A, Impurity B, and/orUnknown C present in the composition. In some embodiments, oxygenpresent in the headspace of a vial containing the composition may bereduced by replacing some or all of the oxygen a gas other than oxygen,preferably an inert gas, thereby at least partially reducing the amountof Impurity A, Impurity B, and/or Unknown C present in the composition.In some embodiments, some or all of the oxygen present in the headspaceof a vial containing the composition may be replaced with nitrogenand/or argon.

In some embodiments, other components of the composition may be presentat a concentration that reduces the amount of Impurity A, Impurity B,and/or Unknown C present in the composition after a certain period ofshelf life and/or reduces the rate of Impurity A, Impurity B, and/orUnknown C formation. In some embodiments, other components of thecomposition may reduce the amount of Impurity A, Impurity B, and/orUnknown C present in the composition after a certain period of shelflife by interfering with an interaction involving other components ofthe composition. In some embodiments, other components of thecomposition may reduce the amount of Impurity A, Impurity B, and/orUnknown C present in the composition after a certain period of shelflife by interfering with an oxidative processes involving epinephrine.In some embodiments, other components of the composition may reduce thepresence of Impurity A, Impurity B, and/or Unknown C in the compositionafter a certain period of shelf life by interacting with a trace metalpresent in the composition. In some embodiments, the transition metalcomplexing agent may reduce the amount of Impurity A, Impurity B, and/orUnknown C present in the composition after a certain period of shelflife.

The present invention provides for compositions that may have anextended shelf life. As used throughout this application, the term“shelf life” refers to the length of time that a product may be storedwithout becoming unfit for medical use. Examples of compositions whichare unfit for medical use include, but are not limited to, compositionswith unacceptably high impurity levels and/or the presence of physicalchanges described herein, such as color change and/or the presence ofinsoluble particles.

In some embodiments, the period of shelf life of the composition may be1 month, preferably 2 months, more preferably 3 months, more preferably4 months, more preferably 5 months, more preferably 6 months, morepreferably 7 months, more preferably 8 months, more preferably 9 months,more preferably 10 months, more preferably 11 months, more 12 months,preferably 13 months, more preferably 14 months, more preferably 15months, more preferably 16 months, more preferably 17 months, morepreferably 18 months, more preferably 19 months, more preferably 20months, more preferably 21 months, more preferably 22 months, morepreferably 23 months, more preferably 24 months, more preferably 25months, more preferably 26 months, more preferably 27 months, morepreferably 28 months, more preferably 29 months, more preferably 30months, more preferably 31 months, more preferably 32 months, morepreferably 33 months, more preferably 34 months, more preferably 35months, and most preferably 36 months. In some embodiments, the periodof shelf life may vary based on product presentation.

In some embodiments, shelf life may be determined by measuring certaincharacteristics of the composition that may indicate that thecomposition is unfit for medical use. In some embodiments, shelf lifemay be determined by measuring the concentration of impurities in thecomposition after storage at 25° C. and 60% relative humidity. In someembodiments, shelf life may be determined by measuring the concentrationof impurities in the composition after storage at 37° C. and 65%relative humidity.

In some embodiments, shelf life may be determined by measuring theconcentration of impurities in the composition using the guidelines asoutlined in the ICH Harmonised Tripartite Guideline: Stability Testingof New Drug Substances and Products Q1A(R2), dated Feb. 6, 2003, thedisclosure of which is incorporated by reference herein in its entirety.

For example, shelf life may be determined for long term, accelerated,and, where appropriate, intermediate storage conditions by measuring theconcentration of impurities after storage in the following conditions,wherein the composition is packaged in a container closure system thatis the same as or simulates the packaging proposed for storage anddistribution.

Minimum time Study Storage condition period covered Long term 25° C. ±2° C./ 12 months 60% RH ± 5% RH or 30° C. ± 2° C./ 65% RH ± 5% RHIntermediate 30° C. ± 2° C./ 6 months 65% RH ± 5% RH Accelerated 40° C.± 2° C./ 6 months 75% RH ± 5% RH

In some embodiments, shelf life may be attributed at least in part tothe concentration of impurities in the composition such that thereduction of impurity concentration and/or rate of impurity formationlengthens the composition's shelf life.

The present invention provides for compositions that may have an initialpH which at least partially influences any of the mechanisms describedherein. In some embodiments, the initial pH of the composition may besuch that the rate of component degradation is minimized. For example,the initial pH of the composition may be such that the rate of formationof D-epinephrine is minimized after a certain period of shelf life. Insome embodiments, the initial pH range of the composition may be in therange of about 3.5 and 4.5, preferably in the range of about 3.6 and4.4, more preferably in the range of about 3.7 and 4.3, more preferablyin the range of about 3.8 and 4.2, more preferably in the range of about3.9 and 4.1, and most preferably about 4.0.

In some embodiments, components of the composition may be selected suchthat the pH of the composition is within a preferred range. For example,in some embodiments, the pH raising agent may be present in thecomposition such that the pH of the composition is higher than what itwould otherwise be without the presence of the pH raising agent.

In some embodiments, the initial pH may be such that the compositionconforms to industry requirements, such as the limit on total acidityimposed by the USP 37-NF 32, S2, monograph.

The present invention also provides for compositions that may resistsignificant pH change after a certain period of shelf life. In someembodiments, the pH range after a certain period of shelf life of thecomposition may be in the range of about 2.5 and 5, preferably in therange of about 2.6 and 4.9, more preferably in the range of about 2.7and 4.8, more preferably in the range of about 2.8 and 4.7, morepreferably in the range of about 2.9 and 4.6, more preferably in therange of about 3.0 and 4.5, more preferably in the range of about 3.1and 4.4, more preferably in the range of about 3.2 and 4.3, morepreferably in the range of about 3.3 and 4.2, more preferably in therange of about 3.4 and 4.1, and most preferably in the range of about3.5 and 4.0.

In some embodiments, components of the composition may be selected suchthat the pH range after a certain period of shelf life of thecomposition is within a preferred range. For example, in someembodiments, the pH raising agent may be present in the composition suchthat the pH range after a certain period of shelf life of thecomposition is higher than what it would otherwise be without thepresence of the pH raising agent.

In some embodiments, the pH range after a certain period of shelf lifeof the composition may be such that the degradation of components of thecomposition is reduced. In some embodiments, the pH range after acertain period of shelf life of the composition may be such that theformation of D-epinephrine is reduced. In some embodiments, the pH rangeafter a certain period of shelf life of the composition may be such thatthe amount of D-epinephrine present in the composition is in a preferredrange described herein.

In some embodiments, the pH range after a certain period of shelf lifemay be such that the degradation of the antioxidant is reduced. Forexample, for compositions containing sulfites, the conversion ofbisulfite to sulfer dioxide may be reduced. The reduction of theconversion of bisulfite to sulfer dioxide may at least in part lead to ahigher proportion of antioxidant present in the composition after acertain period of shelf life compared to compositions outside of thepreferred pH range.

In some embodiments, the pH range of the composition after a certainperiod of shelf life does not change significantly from the initial pHof the composition. In some embodiments, the change in pH range may beno more than about 1 unit different than the initial pH of thecomposition, preferably no more than about 0.9 units, more preferably nomore than about 0.8 units, more preferably no more than about 0.7 units,more preferably no more than about 0.6 units, more preferably no morethan about 0.5 units, more preferably no more than about 0.4 units, morepreferably no more than about 0.3 units, more preferably no more thanabout 0.2 units, most preferably no more than about 0.1 units. In someembodiments, the pH after a certain period of shelf life may not fallbelow 3.5.

In some embodiments, the pH range may be such that the compositionconforms to the limit on total acidity imposed by the USP 37-NF 32, S2,monograph.

The present invention provides for compositions in single-dose and/ormulti-dose formulations. In some embodiments, the composition may becontained in vials. In some embodiments, the vials may comprise clearglass, amber glass, or plastic. In some embodiments, the vials may be inthe range of about 0.1 to 500 mL in volume, preferably in the range ofabout 0.5 to 250 mL, more preferably in the range of about 1 to 100 mL,and most preferably in the range of about 10 to 50 mL. In someembodiments, the composition may exist in a 1 mL vial. In someembodiments, the composition may exist in a 30 mL vial. In someembodiments, the 1 mL vial may be a single-dose formulation. In someembodiments, the 30 mL vial may be a multi-dose formulation. In someembodiments, the same vial may be used for multiple applications of thecomposition for up to about 10 days after initial use, preferably up toabout 15 days, more preferably up to about 30 days, more preferably upto about 45 days, and most preferably up to about 60 days. In someembodiments, the composition may be lyophilized.

In some embodiments, the composition may be contained in anautoinjector.

In some embodiments, the composition may be contained in a pre-filledsyringe.

The present invention also provides for methods of making pharmaceuticalcompositions of the present invention. In some embodiments of carryingout a method of making the present composition, a bulk solution may beproduced as follows: water for injections, whose dissolved oxygencontent is in an acceptable range as measured by a WFI USP/EP O₂ probe,may be added to a manufacturing tank kept under nitrogen pressure. Then,a tonicity regulating agent may be added and mixed. Then, a pH raisingagent may be added to the solution while mixing. Then, a metalcomplexing agent may then be added to the solution while mixing. Anantioxidant may then be added while mixing.

In another container equipped with a stir bar, a pH lowering agent maybe combined with an active agent. The pH lowering agent and the activeagent together may then be stirred using the stir bar in the separatecontainer before the mixture is added to the manufacturing tank undernitrogen.

Then, the separate container may be rinsed with water for injections,whose dissolved oxygen content is in an acceptable range as measured bya WFI USP/EP O₂ probe, using a pipet and added to the manufacturingtank. This rinsing step may be repeated 1 to 3 more times. The solutionmay then mix before the final net weight of the solution is adjustedusing water for injections.

The present invention also provides for methods of treating or reducingthe symptoms associated with a medical condition, comprisingadministering to a subject in need thereof the pharmaceuticalcomposition of the present invention.

Examples of conditions to be treated comprise bronchospasm, sensitivityreactions, cardiac arrhythmias, GI and renal hemorrhage, superficialbleeding, premature labor, hypoglycemia, and cardiogenic, hemorrhagic,and traumatic shock. In some embodiments, the present invention providesfor methods for reducing the symptoms associated with allergic reactions(Type 1), including anaphylaxis, and/or for the induction andmaintenance of mydriasis during intraocular surgery.

Examples of methods of administration comprise subcutaneous,intracameral, intravenous, and intramuscular injection, infusion,intra-arterial administration, intracardiac injection, endotrachealadministration, intraosseous administration, oral inhalation, topicaladministration, and as ophthalmic irrigation.

Unless otherwise noted, all concentrations herein (other than percentageof impurities) are expressed as weight/volume percent (w/v %) of thecomposition.

The present invention is further described by way of the followingnon-limiting Examples that are given for illustrative purposes only.

Examples 1-24 represent example compositions according to the presentinvention.

Example 1

Ingredient Concentration (mM) Epinephrine 1-10 Tonicity regulating agent75-150 Antioxidant 1-10 pH lowering agent 2-11 Transition metalcomplexing agent 0.1-15  Water Fill to final total volume

Example 2

Ingredient Concentration (mM) Epinephrine 4-8 Tonicity regulating agent 90-140 Antioxidant 2-6 pH lowering agent 4-8 Transition metalcomplexing agent 0.2-2  Water Fill to final total volume

Example 3

Ingredient Concentration (mM) Epinephrine 1-10 Sodium Chloride 75-150Sodium Metabisulfite 1-10 Hydrochloric Acid 4% Solution 2-11Tetraethylenepentamine 0.1-15  Water Fill to final total volume

Example 4

Ingredient Concentration (mM) Epinephrine 1-10 Sodium Chloride 75-150Sodium Metabisulfite 1-10 Hydrochloric Acid 4% Solution 2-11 DisodiumEdetate Dihydrate 0.1-15  Water Fill to final total volume

Example 5

Ingredient Concentration (mM) Epinephrine 1-10 Tonicity regulating agent75-150 pH raising agent 10-70  Antioxidant 1-10 pH lowering agent 2-11Water Fill to final total volume

Example 6

Ingredient Concentration (mM) Epinephrine 4-8 Tonicity regulating agent 90-140 pH raising agent 30-50 Antioxidant 2-6 pH lowering agent 4-8Water Fill to final total volume

Example 7

Ingredient Concentration (mM) Epinephrine 3-11 Sodium Chloride 80-160Tartaric Acid 7-21 Sodium Hydroxide 15-30  Sodium Metabisulfite 1-10Hydrochloric Acid 4% Solution 1-10 Water Fill to final total volume

Example 8

Ingredient Concentration (mg/mL) Epinephrine 1-9  Sodium Chloride 1-10Tartaric Acid 2-15 Sodium Hydroxide 1-5  Sodium Metabisulfite 1-12Hydrochloric Acid 4% Solution 0-6 (mL/mL) Water Fill to final totalvolume

Example 9

Ingredient Concentration (mM) Epinephrine 1-15 Sodium Chloride 75-150Lactic Acid 5-25 Sodium Hydroxide 15-30  Sodium Metabisulfite 1-10Hydrochloric Acid 4% Solution 1-10 Water Fill to final total volume

Example 10

Ingredient Concentration (mM) Epinephrine 1-10 Tonicity regulating agent75-150 pH raising agent 10-70  Antioxidant 1-10 pH lowering agent 2-11Transition metal complexing agent 0.1-15  Water Fill to final totalvolume

Example 11

Ingredient Concentration (mM) Epinephrine 4-8 Tonicity regulating agent 90-140 pH raising agent 30-50 Antioxidant 2-6 pH lowering agent 4-8Transition metal complexing agent 0.2-2  Water Fill to final totalvolume

Example 12

Ingredient Concentration (mM) Epinephrine 3-11 Sodium Chloride 80-160Tartaric Acid 7-21 Sodium Hydroxide 15-30  Sodium Metabisulfite 1-10Hydrochloric Acid 4% Solution 1-10 Disodium Edetate Dihydrate 0-5  WaterFill to final total volume

Example 13

Ingredient Concentration (mM) Epinephrine 1-15 Sodium Chloride 75-150Lactic Acid 5-25 Sodium Hydroxide 15-30  Sodium Metabisulfite 1-10Hydrochloric Acid 4% Solution 1-10 Disodium Edetate Dihydrate 0.5-8.5 Water Fill to final total volume

Example 14

Ingredient Concentration (mM) Epinephrine 1-15 Sodium Chloride 50-150Tartaric Acid 5-20 Sodium Hydroxide 15-30  Sodium Metabisulfite 3-18Hydrochloric Acid 4% Solution 1-10 Tetraethylenepentamine 0.5-10.5 WaterFill to final total volume

Example 15

Ingredient Concentration (mM) Epinephrine 1-10 Tonicity regulating agent75-150 pH raising agent 10-70  Antioxidant 1-10 Preservative 10-50  pHlowering agent 2-11 Transition metal complexing agent 0.1-15  Water Fillto final total volume

Example 16

Ingredient Concentration (mM) Epinephrine 4-8 Tonicity regulating agent 90-140 pH raising agent 30-50 Antioxidant 2-6 Preservative 20-40 pHlowering agent 4-8 Transition metal complexing agent 0.2-2  Water Fillto final total volume

Example 17

Ingredient Concentration (mg/mL) Epinephrine 1-10 Sodium Chloride 4-12Lactic Acid 0.5-15   Sodium Hydroxide 1-5  Sodium Metabisulfite 0.1-10.1Chlorobutanol, hydrous 3-12 Hydrochloric Acid 4% Solution 0.1-5 (mL/mL)Disodium Edetate Dihydrate 1-15 Water Fill to final total volume

Example 18

Ingredient Concentration (mM) Epinephrine 1-15 Sodium Chloride 50-150Tartaric Acid 5-20 Sodium Hydroxide 15-30  Sodium Metabisulfite 3-18Chlorobutanol, hydrous 10-30  Hydrochloric Acid 4% Solution 1-10Tetraethylenepentamine 0.5-10.5 Water Fill to final total volume

Example 19

Ingredient Concentration (mM) Epinephrine 0.62 Sodium Chloride 133.47Tartaric Acid 14.99 Sodium Hydroxide 25.00 Glutathione 4.88 HydrochloricAcid 0.69 Disodium Edetate Dihydrate 0.54 Water Fill to final totalvolume

Example 20

Ingredient Concentration (mg/mL) Epinephrine 0.114 Sodium Chloride 7.8Tartaric Acid 2.25 Sodium Hydroxide 1 Glutathione 1.5 Hydrochloric Acid0.025 Disodium Edetate Dihydrate 0.2 Water Fill to final total volume

Example 21

Ingredient Concentration (mM) Epinephrine 0.62 Sodium Chloride 133.47Tartaric Acid 14.99 Sodium Hydroxide 25.00 Methionine 4.89 HydrochloricAcid 0.69 Disodium Edetate Dihydrate 0.54 Water Fill to final totalvolume

Example 22

Ingredient Concentration (mg/mL) Epinephrine 0.114 Sodium Chloride 7.8Tartaric Acid 2.25 Sodium Hydroxide 1 Methionine 0.73 Hydrochloric Acid0.025 Disodium Edetate Dihydrate 0.2 Water Fill to final total volume

Example 23

Ingredient Concentration (mM) Epinephrine 0.62 Sodium Chloride 133.47Tartaric Acid 14.99 Sodium Hydroxide 25.00 Ascorbic Acid 4.83Hydrochloric Acid 0.69 Disodium Edetate Dihydrate 0.54 Water Fill tofinal total volume

Example 24

Ingredient Concentration (mg/mL) Epinephrine 0.114 Sodium Chloride 7.8Tartaric Acid 2.25 Sodium Hydroxide 1 Ascorbic Acid 0.85 HydrochloricAcid 0.025 Disodium Edetate Dihydrate 0.2 Water Fill to final totalvolume

Example 25

The following study was performed in order to compare the stability ofcompositions according to the present invention with otherepinephrine-containing compositions. In particular, a 1 mL formulationaccording to Example 19 and a 1 mL formulation according to ComparativeComposition 1 (below) were subjected to both accelerated (40° C.) andlong term (25° C.) storage conditions for 3 months and 6 months,respectively.

Comparative Composition 1

Ingredient Concentration (mg/mL) Epinephrine 0.114 Sodium Chloride 7.8Tartaric Acid 2.25 Sodium Hydroxide 1 Sodium Metabisulfite 0.457Hydrochloric Acid 0.025 Disodium Edetate Dihydrate 0.2 Water Fill tofinal total volume

The total concentration of impurities was determined at various timepoints during the study. The impurities measured included D-epinephrine,ESA, adrenalone, Impurity A, Impurity B, Unknown C, and adrenochromesulfonate.

The results of this study for both long term and accelerated storageconditions are shown in FIG. 5 . In particular, FIG. 5 shows that thecomposition according to Example 19 surprisingly shows significantlyless impurities after 3 months at both long term and accelerated storageconditions when compared to Comparative Composition 1.

Example 26

In this study, a 1 mL formulation according to Example 19 and a 1 mLformulation according to Comparative Composition 2 (below) weresubjected to accelerated (40° C.) and/or long term (25° C.) storageconditions for 3 months.

Comparative Composition 2

Ingredient Concentration (mg/mL) Epinephrine 0.114 Sodium Chloride 7.8Tartaric Acid 2.25 Sodium Hydroxide 1 Sodium Metabisulfite 0.2285Hydrochloric Acid 0.025 Disodium Edetate Dihydrate 0.2 Water Fill tofinal total volume

The total concentration of impurities was determined at various timepoints during the study. The impurities measured included D-epinephrine,ESA, adrenalone, Impurity A, Impurity B, Unknown C, and adrenochromesulfonate.

The results of this study for both long term and accelerated storageconditions are shown in FIG. 6 . In particular, FIG. 6 shows that thecomposition according to Example 19 surprisingly shows significantlyless impurities after 3 months at accelerated storage conditions whencompared to Comparative Composition 2, and a surprisingly lowconcentration of impurities after 3 months at long term storageconditions.

Example 27

The following is an example procedure for separating and characterizingsubstances that may be present in pharmaceutical compositions comprisingepinephrine. Specifically, this procedure utilizes High PerformanceLiquid Chromatography (HPLC) to separate certain components. Individualcomponents may then be detected and determined by their UV absorbanceresponse at a wavelength of 280 nm.

The chromatographic conditions utilized in this procedure are asfollows:

Instrument Agilent 1100 series Column 4.6 × 150 mm, SeQuant ZIC-HILIC3.5 μm, 100 Å (EMD Millipore Part No. 1.50444.0001) Column Temperature35° C. Flow Rate 1.0 mL/min Injection Volume 100 μL Detector UVAbsorbance with a Diode Array Detector Signal Bandwidth (nm) (nm)Reference 280 10 OFF 292 10 OFF 346 10 OFF 380 10 OFF Run Time 35minutes Pump Time % Mobile % Mobile (Gradient) (minutes) Phase A Phase B0 87 13 12 87 13 22 70 30 25 70 30 28 87 13 35 87 13

The reagents necessary for this procedure are as follows: water, Milli-Qor equivalent; Formic Acid, 88% ACS Reagent Grade; Ammonium Hydroxide,6.0 M Reagent Grade or equivalent; Sodium Chloride, ACS Reagent Grade orequivalent; Acetonitrile, HPLC grade or equivalent; and 0.9% SalineSolution.

From these reagents, the following preparations are prepared:

Ammonium Formate Buffer

First, 1.5 g of Sodium Chloride is dissolved in 1.0 L of water and thenfiltered through a 0.45 μm nylon membrane. Then, 5.0 mL of Formic Acid,88% and 3.0 mL of 6.0 M Ammonium Hydroxide are added and mixedthoroughly. The resulting solution is about 133 mM in Formic Acid and 18mM in Ammonia with a pH about 2.9.

Mobile Phase A

100 mL of the Ammonium Formate Buffer is mixed with 900 mL ofAcetonitrile.

Mobile Phase B

100 mL of the Ammonium Formate Buffer is mixed with 400 mL of water and500 mL of Acetonitrile.

Stock Standard Solution

First, 45 mg of USP Epinephrine Bitartrate Reference Standard or aqualified In-House Reference Standard is accurately weighed andtransferred into a 25 mL amber volumetric flask. This is then dissolvedin and diluted to volume with the Ammonium Formate Buffer. The solutionis mixed by inverting and shaking the volumetric flask at least tentimes.

Intermediate Standard Solution

First, 2.0 mL of the Stock Standard Solution is pipetted into a 200 mLvolumetric flask. The solution is then diluted to volume with theAmmonium Formate Buffer. It is mixed by inverting and shaking thevolumetric flask at least ten times.

Working Standard Preparation

First, 5.0 mL of the Intermediate Standard Solution is pipetted into avolumetric flask. Then, 15.0 mL of Acetonitrile is pipetted into thesame volumetric flask. (The volume is not diluted). The solution ismixed thoroughly. (Standard Preparations stored at room temperatureprotected from light are stable for 48 hours.)

Blank Preparation #1

First, 5.0 mL of the Ammonium Formate Buffer is pipetted into avolumetric flask. Then, 15.0 mL of Acetonitrile is pipetted into thesame volumetric flask. (The volume is not diluted.) The preparation ismixed thoroughly.

Blank Preparation #2

First, 5.0 mL of 0.9% Saline is pipetted into a volumetric flask. Then,15.0 mL of Acetonitrile is pipetted into the same volumetric flask. (Thevolume is not diluted.) The preparation is mixed thoroughly.

Photodegraded Solution

In a clean, clear 2 L glass bottle, 800 mL of water is added. Then,about 6.147 g of NaCl is added followed by about 2.251 g of TartaricAcid. The components are mixed to dissolve. 6.25 mL of 5.0 N NaOH isthen added and mixed well. Then, about 1.425 g of Sodium Metabisulfiteis added. About 1.1334 g of Epinephrine is then diluted with 6.6 mL of1.0 N HCl and quantitatively transferred to the 2 L glass bottle. Thesolution is then diluted to a final volume of 1 L and exposed tocontinuous light in the hood for 12 days, or in a photostability lightchamber for two days.

EpiHILIC IMS (Impurity Marker Solution)

500 mL of the photodegraded solution is added to a 500 mL clear glassbottle. Then, 6.0 mL of 1.0 N HCl is added and mixed well. About 9.5 mgof ESA is added and mixed to dissolve. About 5.6 mg of Adrenochrome isthen added and mixed to dissolve. Then, exactly 10 mL of this solutionis withdrawn and about 4.7 mg of Adrenalone is added. It is mixed todissolve. 0.8 mL of the Adrenalone stock is then added back to the bulkmixture and mixed to dissolve. The remaining Adrenalone stock isdiscarded. The solution is mixed well and transferred to individualamber Autosampler vials. The vials are stored refrigerated or frozen.

Intermediate IMS

5.0 mL of the EpiHILIC IMS is pipetted into a 50 mL volumetric flask anddiluted to volume with 0.9% Saline Solution.

Working IMS

One volume of the Intermediate IMS is diluted with about 3 volumes ofAcetonitrile.

Test samples may then be injected. The Working Standard Preparationsshould be injected after every 4 sample injections or less and thebracketing standard peak areas should be averaged for use in thecalculations. The chromatograms and the Epinephrine and RelatedSubstance peak areas should be measured using the outlined parameters.The Epinephrine Bitartrate peak area in each ongoing standard must be90.0-110.0% of the average area of Epinephrine Bitartrate in n=5injections of Working Standard Preparation injections.

After the completion of each chromatographic run, the column is washedas follows: 20 column volumes of deionized water; 30 column volumes of0.5M NaCl; 30 column volumes of deionized water; 20 column volumes ofacetonitrile/water (50:50, v/v); 20 column volumes of acetonitrile; and20 column volumes of acetonitrile/water (50:50, v/v).

For identification, the retention time of the main peak in thechromatogram of the Sample Preparation (t_(sample)) is compared with theprior injection of the Working Standard Preparation (t_(std)). For apositive test, Rr should be 0.98 to 1.02, wherein Rr is(t_(sample)/t_(std)).

The following information can then be calculated:

Assay(mg/mL)=(A _(E) /A _(s))×(S/25)×(2/200)×(P/100)×(183.20/333.29)

Assay (% LabelClaim)=(Assay (mg/mL)/0.1 nm/mL)×100%

% Related Substances(RS)=(A _(RS) /A_(s))×RRM×(S/25)×(2/200)×(P/100)×(183.20/333.29)×(100%/0.1 mg/ml)

where A_(E) is the Epinephrine peak area for the Sample Preparation;A_(s) is the average Epinephrine peak area for the Bracketing WorkingStandard injections; S is the weight in milligrams of EpinephrineBitartrate in the Stock Standard Preparation; P is the Purity ofEpinephrine Bitartrate in the Stock Standard Preparation expressed as apercentage; 183.20 is the molecular weight of Epinephrine; 333.29 is themolecular weight of Epinephrine Bitartrate; 0.1 mg/mL is the Label Claimfor the Adrenalin® syringe; A_(RS) is the peak area for relatedsubstance in the Sample Preparation integrated at the detectionwavelength in the below table; A_(s) is the average Epinephrine peakarea for the Bracketing Standard injections, and RRF is the RelativeResponse Multiplier from the below table. Specifically, the below tableshows the Relative Retention Time (that is, the time it takes for eachindividual substances to elute from the HPLC column, or RRT) and RRM forseveral specified and unspecified related substances:

Approx. Detection Relative Response Substance RRT Wavelength Multiplier(RRM) Epinephrine 1.00 280 Adrenalone 0.62 280 0.29 ESA 1.91 280 1.37Impurity B 0.57 292 1.09 Impurity A 2.06 292 1.09 Adrenochrome 1.29 3460.35 Sulfonate (AdrS) Unk C 1.84 380 1 Unspecified 280 1

Example 28

The following studies were performed in order to compare the stabilityof certain epinephrine-containing compositions. Specifically, 18 samples(Samples A-P) were subjected to accelerated (40° C.) and/or long term(25° C.) storage conditions for up to 6 months. Each of the 18 samplesconsisted of one of the 6 formulations below.

Formulation 1

Concentration Ingredient mg/mL mM Epinephrine 0.114 0.62 Sodium Chloride7.8 133.47 Tartaric Acid 2.25 14.99 Sodium Hydroxide 1 25.00 SodiumMetabisulfite 0.457 4.81 Hydrochloric Acid 0.025 0.69 Disodium EdetateDihydrate 0.2 0.54 Water for Injection QS to 1 N/A mL

Formulation 2

Concentration Ingredient mg/mL mM Epinephrine 0.114 0.62 Sodium Chloride9 154.00 Tartaric Acid 0.225 1.50 Sodium Hydroxide 0.1 2.50 SodiumMetabisulfite 0.0457 0.48 Hydrochloric Acid 0.025 0.69 Disodium EdetateDihydrate 0.02 0.05 Water for Injection QS to 1 N/A mL

Formulation 3

Concentration Ingredient mg/mL mM Epinephrine 0.114 0.62 Sodium Chloride7.8 133.47 Tartaric Acid 2.25 14.99 Sodium Hydroxide 1 25.00 SodiumMetabisulfite 0.2285 2.40 Hydrochloric Acid 0.025 0.69 Disodium EdetateDihydrate 0.2 0.54 Water for Injection QS to 1 N/A mL

Formulation 4

Concentration Ingredient mg/mL mM Epinephrine 0.114 0.62 Sodium Chloride7.8 133.47 Tartaric Acid 2.25 14.99 Sodium Hydroxide 1 25.00 AscorbicAcid 0.85 4.83 Hydrochloric Acid 0.025 0.69 Disodium Edetate Dihydrate0.2 0.54 Water for Injection QS to 1 N/A mL

Formulation 5

Concentration Ingredient mg/mL mM Epinephrine 0.114 0.62 Sodium Chloride7.8 133.47 Tartaric Acid 2.25 14.99 Sodium Hydroxide 1 25.00 Glutathione1.5 4.88 Hydrochloric Acid 0.025 0.69 Disodium Edetate Dihydrate 0.20.54 Water for Injection QS to 1 N/A mL

Formulation 6

Concentration Ingredient mg/mL mM Epinephrine 0.114 0.62 Sodium Chloride7.8 133.47 Tartaric Acid 2.25 14.99 Sodium Hydroxide 1 25.00 Methionine0.73 4.89 Hydrochloric Acid 0.025 0.69 Disodium Edetate Dihydrate 0.20.54 Water for Injection QS to 1 N/A mL

Table 8 shows certain physical attributes of the compositions at varioustime points during the study, including pH and color. (In Table 8, “C/C”denotes “clear/colorless.”)

TABLE 8 Physical attributes of epinephrine-containing formulations atvarious temperatures and time points Condition Time Point SampleFormulation (° C.) (Months) Description pH A 1 25 0 C/C 4.32 A 1 25 3C/C 4.23 A 1 25 3 C/C 4.23 A 1 25 6 C/C 4.28 A 1 25 6 C/C 4.25 A 1 40 0C/C 4.32 A 1 40 1 C/C 4.30 A 1 40 2 C/C 4.26 A 1 40 3 C/C 4.24 A 1 40 3C/C 4.22 A 1 40 6 C/C 4.26 A 1 40 6 C/C 4.26 B 2 25 0 C/C 4.33 B 2 25 3C/C 4.25 B 2 25 3 C/C 4.28 B 2 25 6 C/C 4.30 B 2 25 6 C/C 4.31 B 2 40 0C/C 4.33 B 2 40 1 C/C 4.29 B 2 40 2 C/C 4.32 B 2 40 3 C/C 4.34 B 2 40 3C/C 4.32 B 2 40 6 C/C 4.28 B 2 40 6 C/C 4.16 C 3 25 0 C/C 4.34 C 3 40 0C/C 4.34 C 3 40 0.9 C/C 4.30 C 3 40 2 C/C 4.34 C 3 40 3 C/C 4.18 C 3 406 C/C 4.27 D 3 25 0 C/C 4.34 D 3 25 0.9 C/C 4.29 D 3 25 2 C/C 4.32 D 325 3 C/C 4.28 D 3 25 4.5 C/C 4.35 D 3 25 6 C/C 4.36 D 3 40 0 C/C 4.34 D3 40 0.9 C/C 4.27 D 3 40 2 C/C 4.31 D 3 40 3 C/C 4.24 D 3 40 4.5 C/C4.39 D 3 40 6 C/C 4.65 E 3 25 0 C/C 4.34 E 3 25 0.9 C/C 4.33 E 3 25 2C/C 4.34 E 3 25 3 C/C 4.32 E 3 25 4.5 C/C 4.34 E 3 25 6 C/C 4.31 E 3 400 C/C 4.34 E 3 40 0.9 C/C 4.31 E 3 40 2 C/C 4.30 E 3 40 3 C/C 4.26 E 340 4.5 C/C 4.35 E 3 40 6 C/C 4.27 F 3 25 0 C/C 4.34 F 3 25 0.9 C/C 4.37F 3 25 2 C/C 4.37 F 3 25 3 C/C 4.36 F 3 25 4.5 C/C 4.34 F 3 25 6 C/C4.35 F 3 40 0 C/C 4.34 F 3 40 0.9 C/C 4.35 F 3 40 2 C/C 4.36 F 3 40 3C/C 4.32 F 3 40 4.5 C/C 4.41 F 3 40 6 C/C 4.36 G 3 25 0 C/C 4.34 G 3 250.9 C/C 4.35 G 3 25 2 C/C 4.37 G 3 25 3 C/C 4.37 G 3 25 4.5 C/C 4.31 G 325 6 C/C 4.33 G 3 40 0 C/C 4.34 G 3 40 0.9 C/C 4.35 G 3 40 2 C/C 4.36 G3 40 3 C/C 4.33 G 3 40 4.5 C/C 4.44 G 3 40 6 C/C 4.33 H 4 25 0 C/C 4.10H 4 25 0.9 C/C 4.11 H 4 25 2 C/C 4.10 H 4 25 3 Pale Yellow 4.12 H 4 254.5 Yellow 4.10 H 4 25 6 Yellow 4.12 H 4 40 0 C/C 4.10 H 4 40 0.9 C/C4.09 H 4 40 2 Pale Yellow 4.13 H 4 40 3 Yellow 4.04 H 4 40 4.5 Yellow4.09 H 4 40 6 Dark Yellow 4.13 I 5 25 0 C/C 4.11 I 5 25 0.9 C/C 4.11 I 525 2 C/C 4.11 I 5 25 3 C/C 4.12 I 5 25 4.5 C/C 4.12 I 5 25 6 C/C 4.11 I5 40 0 C/C 4.11 I 5 40 0.9 C/C 4.12 I 5 40 2 C/C 4.12 I 5 40 3 C/C 4.07I 5 40 4.5 C/C 4.11 I 5 40 6 C/C 4.11 J 6 25 0 C/C 4.29 J 6 25 0.9 C/C —J 6 40 0 C/C 4.29 J 6 40 0.9 Brown — K 2 25 0 C/C 4.29 K 2 25 1 C/C 4.35K 2 25 2 C/C 4.32 K 2 25 3 C/C 4.43 K 2 40 0 C/C 4.29 K 2 40 1 C/C 4.36K 2 40 2 C/C 4.39 K 2 40 3 C/C 4.47 L 2 25 0 C/C 4.38 L 2 25 1 C/C 4.35L 2 25 2 C/C 4.36 L 2 25 3 C/C 4.39 L 2 40 0 C/C 4.38 L 2 40 1 C/C 4.35L 2 40 2 C/C 4.37 L 2 40 3 C/C 4.51 M 1 25 0 C/C 4.36 M 1 25 1 C/C 4.39M 1 25 2 C/C 4.29 M 1 25 3 C/C 4.25 M 1 25 4.5 C/C 4.32 M 1 25 6 C/C4.30 M 1 40 0 C/C 4.36 M 1 40 1 C/C 4.34 M 1 40 2 C/C 4.30 M 1 40 3 C/C4.26 M 1 40 4.5 C/C 4.29 M 1 40 6 C/C 4.26 N 1 25 0 C/C 4.34 N 1 25 1C/C 4.39 N 1 25 2 C/C 4.15 N 1 25 4.5 C/C 4.26 N 1 25 6 C/C 4.23 N 1 256 C/C 4.22 N 1 40 0 C/C 4.34 N 1 40 1 C/C 4.33 N 1 40 2 C/C 4.17 N 1 403 C/C 4.19 N 1 40 4.5 C/C 4.21 N 1 40 6 C/C 4.16 N 1 40 6 C/C 4.16 O 225 0 C/C 4.35 O 2 25 1 C/C 4.38 O 2 25 2 C/C 4.33 O 2 25 3 C/C — O 2 400 C/C 4.35 O 2 40 1 C/C 4.34 O 2 40 2 C/C 4.26 O 2 40 3 Brown — P 2 25 0C/C 4.33 P 2 25 1 C/C 4.36 P 2 25 2 C/C 4.12 P 2 25 4.5 C/C 4.24 P 2 256 C/C 4.22 P 2 25 6 C/C 4.23 P 2 40 0 C/C 4.33 P 2 40 1 C/C 4.32 P 2 402 C/C 4.06 P 2 40 3 C/C 4.21 P 2 40 4.5 C/C 4.25 P 2 40 6 C/C 4.20 P 240 6 C/C 4.18

Table 9 shows the concentration of D-epinephrine and L-epinephrine inthe samples compared with assay values at various time points during thestudy. In Table 9, “assay” is defined as the sum of D-epinephrine andepinephrine, that is, the concentration of total epinephrine. Inaddition, in Tables 9-12, “ND” denotes “not determined,” that is, avalue that is below the limit of detection by the detection method. Thedata shown in Tables 9-12 were obtained using the HPLC method outlinedin Example 27.

TABLE 9 D-epinephrine and L-epinephrine in epinephrine-containingformulations at various temperatures and time points L- Condition TimePoint D-Epinephrine Epinephrine Assay Sample (° C.) (Months) (%) (mg/mL)(mg/mL) A 25 0 0.32 0.103 0.103 A 25 3 0.28 0.121 0.122 A 25 3 0.270.121 0.122 A 25 6 0.31 0.113 0.114 A 25 6 0.34 0.113 0.114 A 40 0 0.320.103 0.103 A 40 1 0.18 0.118 0.118 A 40 2 0.41 0.114 0.114 A 40 3 0.400.112 0.113 A 40 3 0.39 0.112 0.113 A 40 6 0.61 0.101 0.102 A 40 6 0.600.101 0.102 B 25 0 0.40 0.123 0.123 B 25 3 0.36 0.121 0.122 B 25 3 0.350.120 0.121 B 25 6 0.55 0.116 0.117 B 25 6 0.54 0.116 0.117 B 40 0 0.400.123 0.123 B 40 1 0.49 0.118 0.123 B 40 2 0.81 0.115 0.116 B 40 3 1.050.114 0.116 B 40 3 1.62 0.114 0.116 B 40 6 2.18 0.107 0.110 B 40 6 0.210.109 0.110 C 25 0 0.31 0.121 0.121 C 40 0 0.31 0.121 0.121 C 40 0.90.27 0.117 0.118 C 40 2 0.38 0.119 0.119 C 40 3 0.42 0.114 0.115 C 40 60.54 0.107 0.108 D 25 0 0.31 0.121 0.121 D 25 0.9 0.35 0.120 0.121 D 252 0.31 0.125 0.125 D 25 3 0.30 0.126 0.127 D 25 4.5 0.32 0.119 0.120 D25 6 0.30 0.125 0.126 D 40 0 0.31 0.121 0.121 D 40 0.9 0.42 0.121 0.122D 40 2 0.76 0.123 0.135 D 40 3 0.89 0.139 0.140 D 40 4.5 1.09 0.1150.117 D 40 6 1.70 0.144 0.146 E 25 0 0.31 0.121 0.121 E 25 0.9 0.400.116 0.117 E 25 2 0.26 0.122 0.122 E 25 3 0.27 0.119 0.120 E 25 4.50.31 0.117 0.118 E 25 6 0.24 0.119 0.120 E 40 0 0.31 0.121 0.121 E 400.9 0.38 0.113 0.114 E 40 2 0.41 0.119 0.119 E 40 3 0.44 0.111 0.112 E40 4.5 0.58 0.104 0.105 E 40 6 0.63 0.104 0.105 F 25 0 0.31 0.121 0.121F 25 0.9 0.33 0.116 0.117 F 25 2 0.29 0.122 0.122 F 25 3 0.27 0.1200.121 F 25 4.5 0.31 0.116 0.117 F 25 6 0.25 0.120 0.121 F 40 0 0.310.121 0.121 F 40 0.9 0.37 0.114 0.115 F 40 2 0.39 0.120 0.120 F 40 30.42 0.113 0.114 F 40 4.5 0.54 0.106 0.107 F 40 6 0.55 0.108 0.109 G 250 0.31 0.121 0.121 G 25 0.9 0.33 0.115 0.116 G 25 2 0.29 0.122 0.122 G25 3 0.26 0.119 0.120 G 25 4.5 0.37 0.116 0.117 G 25 6 0.24 0.120 0.121G 40 0 0.31 0.121 0.121 G 40 0.9 0.38 0.113 0.114 G 40 2 0.40 0.1190.119 G 40 3 0.44 0.111 0.112 G 40 4.5 0.59 0.104 0.105 G 40 6 0.580.105 0.106 H 25 0 0.31 0.118 0.118 H 25 0.9 2.09 0.105 0.108 H 25 22.01 0.110 0.112 H 25 3 0.34 0.109 0.110 H 25 4.5 0.46 0.109 0.110 H 256 0.70 0.115 0.116 H 40 0 0.31 0.118 0.118 H 40 0.9 1.06 0.104 0.106 H40 2 0.58 0.109 0.108 H 40 3 0.98 0.102 0.103 H 40 4.5 0.69 0.993 0.100H 40 6 1.29 0.103 0.105 I 25 0 0.33 0.119 0.119 I 25 0.9 0.31 0.1190.120 I 25 2 0.20 0.119 0.119 I 25 3 0.26 0.119 0.120 I 25 4.5 0.360.117 0.118 I 25 6 0.72 0.123 0.123 I 40 0 0.33 0.119 0.119 I 40 0.90.30 0.115 0.116 I 40 2 0.08 0.118 0.118 I 40 3 0.26 0.112 0.113 I 404.5 3.22 0.103 0.107 I 40 6 4.56 0.109 0.109 J 25 0 0.33 0.118 0.118 J25 0.9 0.40 0.116 0.117 J 40 0 0.33 0.118 0.118 J 40 0.9 — — — K 25 00.29 0.121 0.121 K 25 1 0.31 0.118 0.119 K 25 2 0.79 0.118 0.119 K 25 30.98 0.117 0.118 K 40 0 0.29 0.121 0.121 K 40 1 0.47 0.119 0.120 K 40 20.35 0.114 0.115 K 40 3 0.38 0.114 0.115 L 25 0 0.29 0.121 0.121 L 25 10.43 0.120 0.121 L 25 2 0.76 0.119 0.120 L 25 3 1.03 0.117 0.119 L 40 00.29 0.121 0.121 L 40 1 0.45 0.118 0.119 L 40 2 0.36 0.114 0.115 L 40 30.38 0.113 0.114 M 25 0 0.39 0.122 0.122 M 25 1 0.39 0.122 0.122 M 25 20.16 0.120 0.120 M 25 3 0.38 0.116 0.117 M 25 4.5 0.31 0.122 0.122 M 256 0.30 0.119 0.120 M 40 0 0.39 0.122 0.122 M 40 1 0.42 0.119 0.119 M 402 0.23 0.116 0.116 M 40 3 0.55 0.108 0.109 M 40 4.5 0.77 0.105 0.106 M40 6 2.00 0.096 0.098 N 25 0 0.39 0.124 0.124 N 25 1 0.38 0.121 0.121 N25 2 0.15 0.119 0.119 N 25 4.5 0.30 0.123 0.123 N 25 6 0.28 0.118 0.119N 25 6 0.28 0.117 0.118 N 40 0 0.39 0.124 0.124 N 40 1 0.42 0.119 0.119N 40 2 0.60 0.120 0.121 N 40 3 0.47 0.107 0.108 N 40 4.5 0.52 0.1100.111 N 40 6 0.53 0.104 0.105 N 40 6 0.54 0.105 0.106 O 25 0 0.38 0.1230.123 O 25 1 0.40 0.124 0.124 O 25 2 0.18 0.123 0.123 O 25 3 — — — O 400 0.38 0.123 0.123 O 40 1 0.70 0.120 0.121 O 40 2 0.86 0.118 0.119 O 403 — — — P 25 0 0.39 0.124 0.124 P 25 1 0.42 0.124 0.124 P 25 2 0.220.123 0.124 P 25 4.5 0.43 0.124 0.124 P 25 6 0.41 0.120 0.121 P 25 60.41 0.120 0.121 P 40 0 0.39 0.124 0.124 P 40 1 0.68 0.122 0.123 P 40 20.22 0.121 0.121 P 40 3 1.13 0.109 0.111 P 40 4.5 1.60 0.116 0.118 P 406 2.70 0.110 0.113 P 40 6 2.72 0.110 0.113

Table 10 shows the concentrations of ESA and adrenalone in the samplesat various time points during the study.

TABLE 10 ESA and adrenalone in epinephrine-containing formulations atvarious temperatures and time points Condition Time Point AdrenaloneSample (° C.) (Months) ESA (%) (%) A 25 0 <0.10 <0.10 A 25 3 1.29 ND A25 3 1.32 ND A 25 6 2.73 <0.10 A 25 6 2.61 <0.10 A 40 0 <0.10 <0.10 A 401 3.15 <0.10 A 40 2 5.16 ND A 40 3 8.84 ND A 40 3 8.77 <0.10 A 40 616.52 0.11 A 40 6 16.59 0.12 B 25 0 <0.10 ND B 25 3 0.83 ND B 25 3 ND NDB 25 6 1.77 ND B 25 6 1.68 <0.10 B 40 0 <0.10 ND B 40 1 1.94 <0.10 B 402 2.98 ND B 40 3 5.14 ND B 40 3 5.23 ND B 40 6 9.67 <0.10 B 40 6 9.71<0.10 C 25 0 ND ND C 40 0 ND ND C 40 0.9 2.24 ND C 40 2 5.05 <0.10 C 403 7.88 ND C 40 6 15.98 ND D 25 0 ND ND D 25 0.9 0.32 ND D 25 2 0.76<0.10 D 25 3 1.15 ND D 25 4.5 1.93 <0.10 D 25 6 2.45 ND D 40 0 ND ND D40 0.9 2.43 ND D 40 2 6.21 0.10 D 40 3 10.05 ND D 40 4.5 13.96 0.11 D 406 22.74 ND E 25 0 ND ND E 25 0.9 0.28 ND E 25 2 0.65 <0.10 E 25 3 0.98ND E 25 4.5 1.56 <0.10 E 25 6 2.07 <0.10 E 40 0 ND ND E 40 0.9 2.14 ND E40 2 5.00 <0.10 E 40 3 7.42 <0.10 E 40 4.5 10.76 0.14 E 40 6 14.43 ND F25 0 ND ND F 25 0.9 0.13 ND F 25 2 0.67 <0.10 F 25 3 ND ND F 25 4.5 1.64<0.10 F 25 6 2.05 ND F 40 0 ND ND F 40 0.9 2.21 ND F 40 2 5.00 <0.10 F40 3 7.55 ND F 40 4.5 11.06 <0.10 F 40 6 15.02 ND G 25 0 ND ND G 25 0.90.32 ND G 25 2 0.68 <0.10 G 25 3 1.03 ND G 25 4.5 1.45 <0.10 G 25 6 2.14<0.10 G 40 0 ND ND G 40 0.9 2.20 ND G 40 2 4.97 <0.10 G 40 3 7.33 ND G40 4.5 10.60 0.12 G 40 6 14.71 ND H 25 0 ND ND H 25 0.9 0.41 ND H 25 2ND 10.26 H 25 3 ND ND H 25 4.5 ND ND H 25 6 ND ND H 40 0 ND ND H 40 0.90.55 ND H 40 2 0.14 6.27 H 40 3 0.69 ND H 40 4.5 ND ND H 40 6 0.22 ND I25 0 ND ND I 25 0.9 ND ND I 25 2 ND ND I 25 3 ND ND I 25 4.5 ND <0.10 I25 6 ND ND I 40 0 ND ND I 40 0.9 ND ND I 40 2 ND <0.10 I 40 3 ND 0.15 I40 4.5 ND ND I 40 6 ND ND J 25 0 ND ND J 25 0.9 ND ND J 40 0 ND ND J 400.9 — — K 25 0 ND ND K 25 1 ND ND K 25 2 0.48 <0.10 K 25 3 0.74 ND K 400 ND ND K 40 1 1.80 ND K 40 2 3.56 ND K 40 3 5.29 <0.10 L 25 0 ND ND L25 1 ND ND L 25 2 0.48 ND L 25 3 0.79 ND L 40 0 ND ND L 40 1 1.67 ND L40 2 <0.10 <0.10 L 40 3 5.32 ND M 25 0 ND ND M 25 1 0.38 <0.10 M 25 20.69 <0.10 M 25 3 1.15 ND M 25 4.5 1.72 <0.10 M 25 6 2.34 ND M 40 0 NDND M 40 1 2.10 <0.10 M 40 2 5.53 <0.10 M 40 3 8.30 ND M 40 4.5 13.162.85 M 40 6 15.83 0.16 N 25 0 ND <0.10 N 25 1 0.34 <0.10 N 25 2 0.77<0.10 N 25 4.5 1.67 <0.10 N 25 6 2.28 ND N 25 6 2.35 ND N 40 0 ND <0.10N 40 1 2.15 <0.10 N 40 2 3.46 <0.10 N 40 3 8.41 ND N 40 4.5 12.57 0.10 N40 6 16.45 0.12 N 40 6 16.70 0.13 O 25 0 ND ND O 25 1 0.17 <0.10 O 25 20.43 ND O 25 3 — — O 40 0 ND ND O 40 1 1.14 ND O 40 2 2.44 <0.10 O 40 3— — P 25 0 ND ND P 25 1 0.18 ND P 25 2 0.45 ND P 25 4.5 1.03 ND P 25 61.48 ND P 25 6 1.48 ND P 40 0 ND ND P 40 1 1.31 <0.10 P 40 2 3.29 <0.10P 40 3 5.04 ND P 40 4.5 7.43 <0.10 P 40 6 9.94 ND P 40 6 9.84 ND

Table 11 shows the concentrations of Impurity A, Impurity B, and UnknownC in the samples at various time points during the study.

TABLE 11 Impurity A, Impurity B, and Unknown C in epinephrine-containingformulations at various temperatures and time points Time Point ImpurityA Impurity B Unknown C Sample Condition (° C.) (Months) (%) (%) (%) A 250 0.14 ND 0.28 A 25 3 ND 0.16 0.54 A 25 3 ND 0.15 ND A 25 6 0.24 0.140.69 A 25 6 0.25 0.16 0.78 A 40 0 0.14 ND 0.28 A 40 1 0.68 0.44 1.30 A40 2 0.28 0.23 0.88 A 40 3 ND 0.29 1.05 A 40 3 0.29 0.27 1.06 A 40 60.46 0.33 1.29 A 40 6 0.33 0.36 1.22 B 25 0 <0.10   <0.10   ND B 25 3 ND0.12 ND B 25 3 ND 0.10 ND B 25 6 <0.10   0.10 ND B 25 6 0.24 0.13<0.10   B 40 0 <0.10   <0.10   ND B 40 1 0.35 0.28 ND B 40 2 0.37 0.28ND B 40 3 0.54 0.44 ND B 40 3 0.40 0.41 ND B 40 6 0.86 0.59 <0.10   B 406 0.85 0.65 ND C 25 0 <0.10   <0.10   ND C 40 0 <0.10   <0.10   ND C 400.9 0.50 0.36 0.40 C 40 2 0.34 0.29 0.37 C 40 3 0.44 0.37 0.48 C 40 60.94 0.54 <0.10   D 25 0 <0.10   <0.10   ND D 25 0.9 0.45 0.32 0.25 D 252 1.17 0.86 0.24 D 25 3 0.74 0.60 ND D 25 4.5 1.39 1.01 0.37 D 25 6 2.862.01 0.19 D 40 0 <0.10   <0.10   ND D 40 0.9 0.98 0.68 0.39 D 40 2 4.963.51 1.06 D 40 3 4.85 3.71 1.12 D 40 4.5 4.87 3.63 1.01 D 40 6 6.09 4.871.21 E 25 0 <0.10   <0.10   ND E 25 0.9 0.30 0.21 0.29 E 25 2 0.18 0.150.28 E 25 3 ND 0.20 ND E 25 4.5 0.34 0.49 0.46 E 25 6 0.49 0.17 0.39 E40 0 <0.10   <0.10   ND E 40 0.9 0.58 0.43 0.55 E 40 2 0.63 0.48 0.64 E40 3 0.82 0.97 0.76 E 40 4.5 1.54 1.02 1.01 E 40 6 2.16 1.98 1.20 F 25 0<0.10   <0.10   ND F 25 0.9 0.13 0.13 0.18 F 25 2 0.11 <0.10   0.18 F 253 ND <0.10   ND F 25 4.5 0.16 <0.10   0.20 F 25 6 0.33 0.17 0.13 F 40 0<0.10   <0.10   ND F 40 0.9 0.39 0.30 0.35 F 40 2 0.40 0.33 0.41 F 40 3ND 0.37 0.40 F 40 4.5 0.86 0.59 0.53 F 40 6 1.04 0.91 0.68 G 25 0<0.10   <0.10   ND G 25 0.9 0.33 0.23 0.26 G 25 2 0.14 0.13 0.20 G 25 3ND 0.17 ND G 25 4.5 0.26 0.11 0.41 G 25 6 0.47 0.35 0.34 G 40 0 <0.10  <0.10   ND G 40 0.9 0.46 0.36 0.41 G 40 2 0.76 0.51 0.68 G 40 3 0.950.78 0.74 G 40 4.5 1.57 1.10 0.89 G 40 6 1.76 1.56 0.91 H 25 0 ND ND NDH 25 0.9 ND 13.22  ND H 25 2 0.39 ND ND H 25 3 ND 52.54  ND H 25 4.50.41 50.23  ND H 25 6 0.25 54.14  <0.10   H 40 0 ND ND ND H 40 0.9<0.10   30.78  ND H 40 2 <0.10   ND ND H 40 3 0.18 22.23  ND H 40 4.50.63 14.70  ND H 40 6 <0.10   12.01  ND I 25 0 ND ND ND I 25 0.9 ND NDND I 25 2 ND ND ND I 25 3 ND ND ND I 25 4.5 ND <0.10   ND I 25 6 ND ND<0.10   I 40 0 ND ND ND I 40 0.9 ND ND ND I 40 2 ND ND ND I 40 3 ND<0.10   ND I 40 4.5 ND 0.77 <0.10   I 40 6 ND ND ND J 25 0 ND ND ND J 250.9 ND ND ND J 40 0 ND ND ND J 40 0.9 — — — K 25 0 ND ND ND K 25 1 ND0.13 ND K 25 2 ND ND ND K 25 3 0.31 0.24 <0.10   K 40 0 ND ND ND K 40 10.26 0.23 ND K 40 2 0.44 0.24 ND K 40 3 0.57 0.47 ND L 25 0 ND ND ND L25 1 ND 0.11 ND L 25 2 0.11 <0.10   <0.10   L 25 3 0.14 <0.10   <0.10  L 40 0 ND ND ND L 40 1 ND 0.27 ND L 40 2 0.62 0.55 ND L 40 3 0.71 0.56ND M 25 0 ND <0.10   0.12 M 25 1 0.15 0.12 0.35 M 25 2 0.60 0.41 0.95 M25 3 0.50 0.37 0.45 M 25 4.5 0.34 0.25 0.58 M 25 6 0.55 0.44 ND M 40 0ND <0.10   0.12 M 40 1 0.41 0.32 0.74 M 40 2 0.94 0.71 1.04 M 40 3 1.050.83 1.09 M 40 4.5 2.28 1.59 1.59 M 40 6 4.22 3.50 1.64 N 25 0 ND ND NDN 25 1 <0.10   <0.10   0.22 N 25 2 0.78 0.55 1.46 N 25 4.5 0.13 0.110.54 N 25 6 ND 0.18 0.57 N 25 6 ND 0.14 0.69 N 40 0 ND ND ND N 40 1 0.360.26 0.80 N 40 2 0.47 0.34 ND N 40 3 0.42 0.31 1.18 N 40 4.5 0.42 0.321.56 N 40 6 ND 0.35 1.65 N 40 6 ND 0.38 1.73 O 25 0 <0.10   <0.10   ND O25 1 0.17 0.13 ND O 25 2 0.21 0.20 ND O 25 3 — — — O 40 0 <0.10  <0.10   ND O 40 1 0.79 0.65 ND O 40 2 1.75 1.31 ND O 40 3 — — — P 25 0ND <0.10   ND P 25 1 <0.10   <0.10   ND P 25 2 0.13 0.10 ND P 25 4.50.13 0.12 ND P 25 6 ND 0.14 ND P 25 6 ND <0.10   ND P 40 0 ND <0.10   NDP 40 1 0.27 0.22 ND P 40 2 0.44 0.34 <0.10   P 40 3 0.57 0.40 <0.10   P40 4.5 0.73 0.57 ND P 40 6 0.84 0.67 ND P 40 6 0.89 0.70 ND

Table 12 shows the concentrations of adrenochrome sulfonate and totalimpurities in the samples at various time points during the study.

TABLE 12 Adrenochrome sulfonate and total impurities inepinephrine-containing formulations at various temperatures and timepoints Total Condition Time Point Adrenochrome Impurities Sample (° C.)(Months) Sulfonate (%) (%) A 25 0 ND 0.42 A 25 3 ND 2.27 A 25 3 ND 1.74A 25 6 <0.10 4.11 A 25 6 ND 4.14 A 40 0 ND 0.42 A 40 1 <0.10 5.75 A 40 2ND 7.30 A 40 3 ND 10.58 A 40 3 ND 10.78 A 40 6 ND 19.55 A 40 6 ND 19.41B 25 0 ND 0.40 B 25 3 ND 1.31 B 25 3 ND 0.45 B 25 6 ND 2.42 B 25 6 ND2.59 B 40 0 ND 0.40 B 40 1 ND 3.06 B 40 2 ND 4.44 B 40 3 ND 7.17 B 40 3ND 7.66 B 40 6 ND 13.30 B 40 6 ND 11.42 C 25 0 ND 0.31 C 40 0 ND 0.31 C40 0.9 ND 3.91 C 40 2 ND 6.63 C 40 3 ND 9.59 C 40 6 ND 18.14 D 25 0 ND0.31 D 25 0.9 ND 2.82 D 25 2 ND 3.46 D 25 3 ND 2.79 D 25 4.5 ND 5.02 D25 6 ND 7.81 D 40 0 ND 0.31 D 40 0.9   0.26 5.57 D 40 2 ND 17.56 D 40 3ND 20.87 D 40 4.5   0.72 27.35 D 40 6   0.86 41.90 E 25 0 ND 0.31 E 250.9 ND 1.58 E 25 2 ND 1.65 E 25 3 ND 1.45 E 25 4.5 ND 3.16 E 25 6 ND3.63 E 40 0 ND 0.31 E 40 0.9 ND 4.39 E 40 2 ND 7.65 E 40 3 ND 10.41 E 404.5 ND 14.77 E 40 6 <0.10 20.80 F 25 0 ND 0.31 F 25 0.9 ND 1.04 F 25 2ND 2.63 F 25 3 ND 0.27 F 25 4.5 ND 2.31 F 25 6 ND 2.93 F 40 0 ND 0.31 F40 0.9 ND 3.71 F 40 2 ND 6.91 F 40 3 ND 8.74 F 40 4.5 ND 13.79 F 40 6 ND18.40 G 25 0 ND 0.31 G 25 0.9 ND 1.57 G 25 2 ND 1.61 G 25 3 ND 1.46 G 254.5 ND 2.74 G 25 6 <0.10 3.54 G 40 0 ND 0.31 G 40 0.9 <0.10 3.97 G 40 2ND 7.87 G 40 3 ND 10.24 G 40 4.5 ND 15.17 G 40 6 ND 19.74 H 25 0 ND 0.79H 25 0.9 ND 52.50 H 25 2 ND 65.92 H 25 3 ND 108.68 H 25 4.5 ND 105.96 H25 6 ND 114.33 H 40 0 ND 0.79 H 40 0.9 ND 62.20 H 40 2 ND 32.07 H 40 3ND 72.91 H 40 4.5 ND 75.23 H 40 6 ND 89.39 I 25 0 ND 0.33 I 25 0.9 ND0.31 I 25 2 ND 0.20 I 25 3 ND 0.26 I 25 4.5 ND 0.36 I 25 6 ND 0.92 I 400 ND 0.33 I 40 0.9 ND 1.34 I 40 2 ND 0.48 I 40 3 ND 0.54 I 40 4.5 ND4.40 I 40 6 ND 6.98 J 25 0 ND 0.33 J 25 0.9 ND 2.12 J 40 0 ND 0.33 J 400.9 — — K 25 0 ND 0.29 K 25 1 ND 0.44 K 25 2 ND 1.27 K 25 3 ND 2.27 K 400 ND 0.29 K 40 1 ND 2.76 K 40 2 <0.10 4.59 K 40 3 ND 7.68 L 25 0 ND 0.29L 25 1 ND 0.54 L 25 2 ND 1.35 L 25 3 <0.10 1.96 L 40 0 ND 0.29 L 40 1 ND2.39 L 40 2 ND 5.04 L 40 3 <0.10 7.07 M 25 0 ND 0.51 M 25 1 ND 1.49 M 252 ND 2.81 M 25 3 ND 2.95 M 25 4.5 ND 3.49 M 25 6 ND 3.63 M 40 0 ND 0.51M 40 1 ND 4.15 M 40 2 <0.10 8.79 M 40 3 ND 12.37 M 40 4.5 ND 12.73 M 406 ND 27.86 N 25 0 ND 0.39 N 25 1 ND 1.07 N 25 2 <0.10 3.71 N 25 4.5 ND4.37 N 25 6 ND 3.31 N 25 6 ND 3.46 N 40 0 ND 0.39 N 40 1 ND 4.13 N 40 2ND 4.87 N 40 3 ND 11.34 N 40 4.5 ND 16.21 N 40 6 ND 19.10 N 40 6 ND19.48 O 25 0 ND 0.38 O 25 1 ND 0.87 O 25 2 ND 1.02 O 25 3 — — O 40 0 ND0.38 O 40 1 ND 3.62 O 40 2 <0.10 6.55 O 40 3 — — P 25 0 ND 0.39 P 25 1ND 0.60 P 25 2 ND 0.90 P 25 4.5 ND 1.83 P 25 6 ND 2.03 P 25 6 ND 1.89 P40 0 ND 0.39 P 40 1 ND 2.60 P 40 2 ND 4.29 P 40 3 ND 7.14 P 40 4.5 ND11.22 P 40 6 ND 14.15 P 40 6 ND 14.15

1-19. (canceled)
 20. A pre-filled syringe containing a composition, thecomposition comprising: epinephrine and/or salts thereof, a tonicityregulating agent, a pH raising agent, an antioxidant comprisingglutathione, a pH lowering agent, a transition metal complexing agent,and water.
 21. The pre-filled syringe of claim 20, wherein the tonicityregulating agent comprises sodium chloride.
 22. The pre-filled syringeof claim 20, wherein the pH raising agent comprises tartaric acid andsodium hydroxide.
 23. The pre-filled syringe of claim 20, wherein the pHlowering agent comprises hydrochloric acid.
 24. The pre-filled syringeof claim 20, wherein the transition metal complexing agent comprisesEDTA.
 25. The pre-filled syringe according to claim 20, wherein thecomposition comprises about 20% or less total impurities after 1 monthof storage in accelerated storage conditions.
 26. The pre-filled syringeaccording to claim 20, wherein the composition comprises about 20% orless total impurities after 2 months of storage in accelerated storageconditions.
 27. The pre-filled syringe according to claim 20, whereinthe composition comprises about 20% or less total impurities after 3months of storage in accelerated storage conditions.
 28. The pre-filledsyringe according to claim 20, wherein the composition comprises about10% or less total impurities after 1 month of storage in acceleratedstorage conditions.
 29. The pre-filled syringe according to claim 20,wherein the composition comprises about 10% or less total impuritiesafter 2 months of storage in accelerated storage conditions.
 30. Thepre-filled syringe according to claim 20, wherein the compositioncomprises about 10% or less total impurities after 3 months of storagein accelerated storage conditions.
 31. A pre-filled syringe containing acomposition, the composition comprising: epinephrine and/or saltsthereof, a tonicity regulating agent comprising sodium chloride, a pHraising agent comprising tartaric acid and sodium hydroxide, anantioxidant comprising glutathione, a pH lowering agent comprisinghydrochloric acid, a transition metal complexing agent comprising EDTA,and water.
 32. The pre-filled syringe according to claim 31, wherein thecomposition comprises about 20% or less total impurities after 1 monthof storage in accelerated storage conditions.
 33. The pre-filled syringeaccording to claim 31, wherein the composition comprises about 20% orless total impurities after 2 months of storage in accelerated storageconditions.
 34. The pre-filled syringe according to claim 31, whereinthe composition comprises about 20% or less total impurities after 3months of storage in accelerated storage conditions.
 35. The pre-filledsyringe according to claim 31, wherein the composition comprises about10% or less total impurities after 1 month of storage in acceleratedstorage conditions.
 36. The pre-filled syringe according to claim 31,wherein the composition comprises about 10% or less total impuritiesafter 2 months of storage in accelerated storage conditions.
 37. Thepre-filled syringe according to claim 31, wherein the compositioncomprises about 10% or less total impurities after 3 months of storagein accelerated storage conditions.
 38. The pre-filled syringe accordingto claim 31, wherein the antioxidant consists of glutathione.
 39. Apre-filled syringe containing a composition, the composition comprising:0.62 mM epinephrine and/or salts thereof, 133.47 mM sodium chloride,19.99 mM tartaric acid, 15 mM sodium hydroxide, 4.88 mM glutathione,0.69 mM hydrochloric acid, 0.54 mM EDTA, and water.